Mammalian ZPAs

ABSTRACT

A method for specifically inducing transient infertility or permanent sterility in a host animal by selective vaccination with specific zona pellucida proteins or immunocontraceptively active fragments thereof. Novel zona pellucida DNA sequences encoding specific zona pellucida proteins are disclosed.

This is a divisional of U.S. application Ser. No. 08/149,223 filed Nov.9, 1993 and currently pending; which is a continuation-in-part of U.S.application Ser. No. 08/012,990 filed Jan. 29, 1993, now abandoned;which is a continuation-in-part of U.S. application Ser. No. 07/973,341filed Nov. 9, 1992, now abandoned.

FIELD OF THE INVENTION

This invention relates generally to the production and use of zonapellucida proteins, and more particularly to novel DNA sequencesencoding zona pellucida proteins, to recombinant materials and methodsfor producing such proteins and to materials and methods for selectivelyeffecting either transient infertility or permanent sterility in mammalsthrough use of naturally occurring and recombinant zona pellucidaproteins.

BACKGROUND OF THE INVENTION

The present invention relates to a method for inducing reproducibletransient infertility or sterility in a mammal by inducing in thatmammal antibodies directed to proteins found in the zona pellucida ofthat mammal's oocytes. The invention also relates to purified, isolatedDNA sequences encoding the zona pellucida proteins herein designated"ZPA" and "ZPB" and "ZPC" from various mammalian species. The inventionis further directed to pharmaceutical compositions capable of inducingantibody production in a subject mammal.

The zona pellucida (ZP) is a complex matrix surrounding the mammalianoocyte, formed of glycoproteins secreted by ovarian cells. Zonapellucida glycoproteins perform a variety of functions. For example, themouse ZP proteins previously designated ZP2 and ZP3 are complexed intolong filaments which are cross-linked by the protein designated ZP1 inthe ZP matrix providing structural integrity to the matrix. Wassarman,P. M., Annu. Rev. Biochem. 57:415-442 (1988). In addition to itsstructural role, mouse ZP3 has been shown to be a sperm receptor in theZP matrix. Bleil, J. P. and Wassarman, P. M., Cell 20: 873-882 (1980).Following binding of sperm to ZP3 and the subsequent induction of thesperm acrosome reaction on the surface of the ZP, ZP2 acts as asecondary sperm receptor that is necessary for the maintenance of spermbinding to the egg. Bleil et al., Dev. Biol. 128: 376-385 (1988).Because of its role in the maintenance of the oocyte and in sperm-oocyteinteractions, the ZP represents a logical target for design ofcontraceptive agents which interfere with the fertilization process.

Various groups have undertaken an immunological approach in attempts tointerfere with ZP functions and thus to decrease fertility in immunizedanimals. See, Dunbar et al. In: International Congress on ReproductiveImmunology. T. Wegman and T. Gills (eds.). London: Oxford Press, pp.505-528 (1983); and Dunbar et al. In: Mechanisms and Control of AnimalFertilization. J. Hartman (ed.) Academic Press, New York, pp. 139-166(1983). These studies showed that active immunization of mammals withovarian homogenates decreased fertility. However, the large number ofcomponents in such homogenates made the identification of antigensresponsible for the decrease in fertility nearly impossible. Inaddition, the use of such a complex mixture creates a potential forunwanted and potentially harmful side-effects.

Research by various investigators using chromatographic methodsincluding SDS polyacrylamide gel electrophoresis (PAGE) and highpressure liquid chromatography (HPLC) have resulted in theidentification of numerous zona pellucida proteins from a variety ofmammalian species. Data compiled by Timmons and Dunbar in "Perspectivesin Immunoreproduction: Conception and Contraception"; pp. 242-260,Mathur, S. and Fredericks, C. M. eds.; New York, Hemisphere PublishingCo (1988), as described below, illustrate examples of zona pellucidaproteins that have been characterized.

Zona pellucida proteins isolated from pig include: PZI, a 40-110 kDprotein isolated by Dunbar et al., Biol. Reprod. 24:1111 (1981); PZII, a70-110 kD protein, PZIII, a 95-118 kD protein, and PZIV, an 18-25 kDprotein, all isolated by Dunbar et al., Biol. Reprod. 32:619 (1985);90K, a 89-119 kD protein, 65K, a 61-83 kD protein, 55K, a 47-66 kDprotein, and 25K, an 18-26 kD protein, all isolated by Hedrick, J. L.and Wardrip, N. J. Biochem. 157: 63 (1986); ZP1, an 82-118 kD protein,ZP2, a 58-96 kD protein, ZP3 (PPZA), a 40-74 kD protein, and ZP4, a 21kD protein, all isolated by Subramanian et al., Biol. Reprod. 24:933(1981); 87K (ZP1I/ZP2), a 77-97 kD protein, 58K, a 40-70 kD protein bothisolated by Yurewicz et al., Biol. Reprod. 29: 511 (1983);deglycosylated PZI, a 35 kD protein; PZII, a 55 kD protein; and PZIII,an 80 kD protein all isolated by Skinner and Dunbar as described inImmunological Approaches to Contraception and the Promotion ofFertility, G. P. Talwar (ed.) New York: Plenum pp. 251-268 (1986); anddeglycosylated ZP3 having a molecular weight of 45 kD isolated by Saccoet al., J. Reprod. Fertil. 76:575 (1986).

Isolated rabbit zona pellucida proteins include: RZI, RZII, and RZIII,having molecular weights of 68-125 kD, 80-100.5 kD, and 100-132 kDrespectively, all isolated by Dunbar et al., Biol. Reprod. 24:1111(1986); ZP1, ZP2, and ZP3 having molecular weights of 100-118 kD, 83-110kD, and 80-92 kD respectively, all isolated by Sacco et al., Proc. Soc.Exp. Biol. Med. 167:318 (1981); deglycosylated RZI, and RZII havingmolecular weights of 65 kD, and 80 kD respectively, both isolated bySkinner and Dunbar and described in Immunological Approaches toContraception and Promotion of Fertility. G. P. Talwar (ed.). New York:Plenum, pp. 251-268 (1986); and deglycosylated RZIII, a 90 kD proteinisolated by Timmons and Dunbar, Biol. Reprod. 36: 1275 (1987).

A number of mouse zona pellucida proteins have been isolated including:ZP1, ZP2, and ZP3 having molecular weights of 200 kD, 120 kD, and 83 kDrespectively, all isolated by Bleil and Wassarman Dev. Biol. 76:185(1980); and ZP1 and ZP2 having molecular weights of 166-122 kD and 90-92kD respectively, isolated by Sacco et al., Proc. Soc. Exp. Biol. Med.167: 318 (1981). The differences in the molecular weights of mouse ZP1and ZP2 as reported by Bleil et al. and Sacco et al. may be due to thefact that Bleil used 2D-PAGE under non-reducing conditions while Saccoused 2D-PAGE under reducing conditions.

The cat zona pellucida proteins CZI and CZII were isolated by Maresh andDunbar J. Exp. Zool. 244:299 (1987) and have molecular weights of 50-110kD and 90-110 kD respectively.

Maresh and Dunbar J. Exp. Zool. 244:299 (1987), have also isolated thedog zona pellucida proteins DZI, DZII, and DZIII which have molecularweights of 50-110 kD, 70-95 kD, and 90-100 kD respectively.

Sacco et al., Proc. Soc. Exp. Biol. Med. 167:318 (1981) describedsquirrel monkey ZP1, ZP2, ZP3, and ZP4 having molecular weights of 63-78kD, 63-70 kD, 47-51 kD, and 43-47 kD respectively. In the samepublication Sacco et al. described human ZP1, ZP2, and ZP3 havingmolecular weights of 80-120 kD, 73 kD, and 59-65 kD respectively.

To date, few mammalian zona pellucida genes or proteins have beenisolated and sequenced. None has been successfully used to produce aneffective immunocontraceptive. A lack of consensus among those of skillin the art regarding the number and characteristics (e.g. molecularweight) of proteins present in the zona pellucida of various mammalianspecies, and difficulties in purifying these heavily glycosylatedproteins have hampered attempts to utilize zona pellucida proteins toproduce an effective immunocontraceptive with predictable function.

A number of groups have had success in cloning cDNAs or genes encodingvarious mammalian zona pellucida proteins.

Ringuette et al., Dev. Biol., 127:287-295 (1988) and Liang et al., Mol.Cell. Biol., 10:1507-1515 (1990), reported cloning of mouse DNA encodingzona pellucida proteins ZP3 and ZP2, respectively. The clones wereobtained by screening mouse cDNA libraries with anti-ZP3 and anti-ZP2antibodies. No sequence homology was found between mouse ZP3 and ZP2.

Ringuette et al., Proc. Natl. Acad. Sci. USA, 83:4341-4345 (1986),reported isolation of a partial cDNA clone for mouse ZP3, which clonehybridized with total genomic DNA of mouse, rat, dog, cow, and human,but not with pig or rabbit genomic DNA unless the hybridization wasperformed at very low stringency. The full length ZP3 cDNA characterizedby Ringuette Dev. Biol. 127:287-295(1988) represents a germ-linespecific mRNA having relatively short 5' and 3' untranslated regions andan open reading frame of about 1317 nucleotides with an additional200-300 nucleotide poly-A tail. Ringuette also found that rat, rabbit,dog, and cow ovary transcribes mRNA which hybridized to the mouse ZP3cDNA and that the ZP3 transcripts had similar molecular weights. Lianget al. Mol. Cell. Biol., 10:1507-1515 (1990), showed that the nucleicacid and deduced amino acid sequence of ZP2 is distinctly different fromthat of ZP3 although it had the same short motif of 5' and 3'untranslated regions. The ZP2 mRNA is reported to have single openreading frame of 2,139 nucleotides which codes for a polypeptide of80,217 Daltons representing 713 amino acids.

Chamberlin and Dean, Dev. Biol. 131:207-214 (1989) and Kinloch, R. A. etal., Proc. Nat. Acad. Sci. USA, 85:6409-6413 (1988) have reported thecloning of the mouse ZP3 gene. The mouse ZP3 gene is reported to have 8exons and 7 introns in a transcription unit of 8.6 kbp.

Kinloch et al., Dev. Biol. 142:414-421 (1990), reported cloning ofhamster genomic ZP3 DNA from a hamster genomic DNA library screened withmouse ZP3 DNA as a probe. The hamster ZP3 gene has a transcription unitof 7900 nucleotides and was found to contain 7 introns and 8 exons. Thehamster ZP3 protein is approximately 81% homologous to mouse ZP3protein. The hamster transcript contained 1266 nucleotides, six lessthan mouse ZP3 mRNA.

Chamberlain and Dean, Proc. Natl. Acad. Sci. USA 87:6014-6018 (1990),reported the cloning of human ZP3 from a human genomic DNA library usingmouse ZP3 cDNA as a probe. The human ZP3 gene is composed of 8 exons ina transcription unit of 18.3 kbp. The exons are almost identical in sizeto the eight exons of mouse ZP3 and the nucleotide sequence of thecoding region is 74% homologous. The human ZP3 transcript is verysimilar to mouse ZP3 mRNA. Both have short 5' and 3' untranslatedregions, and both have a single open reading frame of 1272 nucleotidesthat encodes a 424-amino acid protein.

U.S. Pat. No. 4,996,297, to Dunbar, reported the isolation of threerabbit zona pellucida clones encoding rabbit ZP1 and ZP2 proteins, usinganti-ZP1 and anti-ZP2 antibodies as screening probes. The sequencesdesignated as P2 and P3 in FIG. 4 of the Dunbar patent represent rabbitZP cDNAs of 812 and 1705 nucleotides respectively.

Schwoebel et al., J. Biol. Chem. 266:7214-7219 (1991), isolated andcharacterized a full length cDNA (designated rc 55) encoding the 55-kDrabbit zona pellucida protein using cross-species affinity purifiedantisera. The protein encoded by this cDNA has some similarity to themouse ZP2 protein described by Liang. However, comparisons of rc 55 withthe mouse ZP3 protein revealed no homology.

The functional activities of the cloned ZP DNAs and their encodedproteins have not been fully characterized and neither has theirpotential use as immunocontraceptives been demonstrated.

In order to develop a useful zona pellucida product for use in fertilitycontrol, particularly in the form of a vaccine, it is highly desirableto purify, isolate, and characterize zona pellucida proteins from aspecies of an animal of interest. Because of factors such as the purityof such proteins needed for vaccine production, and the high cost andnumerous problems associated with purification of these proteins, itwould be highly desirable to ascertain the DNA and amino acid sequencesof zona pellucida proteins of a specific species of interest. Havingsuch known, isolated and characterized zona pellucida proteins, thefunction of each zona pellucida protein may be understood and afertility control product may be designed based upon the specificfunctional characteristics of a particular zona pellucida protein andfor a particular mammalian species.

It would be thus highly useful and desirable to provide isolated,purified, sequenced, and characterized recombinant zona pellucidaproteins which would permit the development of fertility controlproducts possessing specific reproducible effects in eliciting transientand/or permanent infertility. Such products, where used to elicittransient infertility, would desirably have long lasting effects so asto minimize the number of times the immunocontraceptive agent must beadministered to maintain infertility.

SUMMARY OF THE INVENTION

The present invention provides novel methods and materials for inducingeither reproducible transient or permanent infertility effects in femalemammals, including humans, by selective administration of homologousand/or heterologous mammalian species ZP proteins orimmunocontraceptively active fragments thereof hereinafter designated asZPA, ZPB and ZPC. By "reproducible" is meant that, unlike prior artattempts to induce transient infertility by administration of ZPproteins (in the form of mixtures of such proteins), this inventionachieves its transient infertility effects by the administration of ZPAand/or ZPB in a form such that the duration of transient infertility iscontrollable and can be maintained in an on or off condition in acontrollable and/or predictable fashion. This is achieved primarilythrough administration of the highly pure ZPA and ZPB proteins orimmunocontraceptively active fragments thereof of this invention, e.g.,in recombinant form and thus essentially devoid of ZPC. Byimmunocontraceptively active fragments is meant a ZP protein fragmentcapable of inducing infertility.

In one of its aspects, the present invention provides methods forinducing reproducible transient infertility in a mammal by administeringto a subject female mammal a zona pellucida protein (or fragmentthereof) selected from the group consisting of mammalian ZPA, and ZPB,and combinations thereof in doses effective to stimulate production insaid mammal of antibodies which recognize ZPA or ZPB proteins of saidmammal. It is presently preferred that mammalian ZPA and ZPB for use insuch methods be derived from the same mammalian species as the subjectmammal although the use of heterologous species proteins is alsocontemplated. Use of purified isolates of mammalian ZPA or ZPB proteinsuch as obtained by chromatographic separatory procedures iscontemplated. Use of proteins produced by recombinant methods isexpected to be most preferred.

According to another aspect of the invention, methods are provided forinducing permanent sterility in a female mammal by administering to asubject female mammal a recombinant mammalian ZPC protein (or fragmentthereof) in a form essentially devoid of ZPA and/or ZPB, in a doseeffective to stimulate production in said female mammal of antibodieswhich recognize the ZPC protein of said mammal. As is the case withinduction of transient infertility, use of homologous species ZPC ispreferred, but not required, and the protein may be derived from naturalsources or produced by recombinant methods. Modified ZPC proteinsincluding but not limited to palmitylated and chitosan modified proteinsare also contemplated by the present invention.

Presently preferred ZPA, ZPB, and ZPC proteins for veterinaryapplication of the transient infertility and sterility inducing methodsinclude porcine, rabbit, canine, feline, bovine, and cynomolgus monkeyZP proteins.

In another of its aspects, the present invention provides pharmaceuticalcompositions for use in inducing reproducible transient infertility in afemale mammal (including humans) comprising an effective dose of a zonapellucida protein (or fragment thereof) selected from the groupconsisting of mammalian ZPA, and ZPB (substantially free of ZPC), incombination with one or more pharmaceutically acceptable carriers,diluents and adjuvants. Modified ZPA and ZPB proteins (for example,palmitylated or chitosan modified) are also contemplated by the presentinvention.

According to another aspect of the present invention, novel purified andisolated DNA sequences are provided which encode porcine ZPA, ZPB, andZPC, as illustrated by the DNA sequences set out in SEQ ID NOS. 1, 3,and 5. Also, provided are purified and isolated DNA sequences encoding:rabbit ZPC, as illustrated by the DNA sequence set out in SEQ ID NO. 7;canine ZPA and ZPC, as illustrated by the DNA sequences set out in SEQID NOS. 9 and 11; feline ZPA, ZPB, and ZPC, as illustrated by the DNAsequences set out in SEQ ID NOS. 13, 15, and 17; bovine ZPA, ZPB, andZPC, as illustrated by the DNA sequences set out in SEQ ID NOS. 19, 21,and 23; human ZPA and ZPB as illustrated by sequences set out in SEQ IDNO. 42 and 40, respectively, and as contained as human DNA inserts inlambda phage clones A1 and A4, (ZPA) and as contained in human DNAinserts in lambda phage clones 1-1 and 4-9 (ZPB).

Polynucleotide sequences of the invention are useful for the productionof ZPA, ZPB and ZPC proteins by recombinant methods and as probes forthe isolation of heterologous species polynucleotides encodingcorresponding zona pellucida proteins by hybridization methods.

Also provided by the present invention are novel host cells, especiallyunicellular eucaryotic and procaryotic cells, stably transformed ortransfected with polynucleotides of the invention in a manner allowingexpression of the ZP proteins (or immunologically significant fragmentsthereof) in the host cells. Host cells expressing such ZP products, whengrown in a suitable culture medium, and particularly useful for largescale production processes wherein the desired polypeptide products, inglycosylated or non-glycosylated form are isolated from the cells or themedium in which the cells are grown.

Recombinant polypeptides provided by the invention thus comprise ZPA,ZPB and ZPC, and full equivalents of such zona pellucida proteinsincluding both glycosylated and non-glycosylated forms, variants andimmunologically active fragments thereof which retain substantialbiological activity, i.e., at least one of the biological activities ofthe zona pellucida protein discussed herein, e.g., the ability tostimulate the production of antibodies as discussed herein uponadministration to a mammal. Such immunologically active fragments may bedefined as containing at least one epitope effective to stimulate theproduction of antibodies upon administration to a mammal in accordancewith this invention.

In another aspect of the invention, a method is provided for theisolation of nucleic acid sequences encoding other mammalian ZPA, ZPB,and ZPC proteins by hybridization under stringent conditions ofheterologous species ZPA, ZPB, and/or ZPC probes to cDNA or genomic DNAlibraries, derived from the mammalian species of interest.

More particularly, it is an aspect of the invention to provide a methodfor the isolation of nucleic acid sequences encoding human ZPA and ZPBby hybridization under stringent conditions of sequences encoding ZPAand/or ZPB from heterologous species.

Other aspects and advantages of the present invention will be readilyunderstood upon consideration of the following detailed description ofpresently preferred embodiments thereof, reference being made to theFIGS. wherein:

DESCRIPTION OF THE FIGURES

FIG. 1 is a diagrammatic representation of the plasmid vector pZ90;

FIG. 2 is a diagrammatic representation of the plasmid vector pZ98; and

FIG. 3 is a diagrammatic representation of the plasmid vector pZ156.

FIG. 4 is a diagrammatic representation of the alignment of the Eco R1fragments encoding human ZPB.

FIG. 5 is a diagrammatic representation of the plasmid vector pZ169.

FIG. 6 is a diagrammatic representation of the plasmid vector pZ145.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to mammalian zona pellucida proteinscharacterized in three major classes: ZPA, ZPB, and ZPC. Thisclassification scheme has resulted from repetitive screening of variousmammalian ovarian cDNA libraries and retrieval of clones which encodeproteins showing significant homology in three distinct groups,designated herein as ZPA, ZPB and ZPC. Although similarity is seenbetween DNA sequences encoding ZPA, ZPB, or ZPC between animal species,very little homology is found between the individual species' ZPA, ZPB,and ZPC proteins.

DNA sequences encoding zona pellucida proteins A, B, and C and theirdeduced amino acid sequences for various mammalian species ZPs arepresented in SEQ ID NOS. 1-24. It is understood that the DNA sequence ofa particular animal may vary slightly due to the phenomenon of allelicvariation. Small differences in the precise DNA sequence between animalsor slight errors due to the inefficiency of sequencing procedures are tobe expected. Such variants are included within the scope of the presentinvention.

The zona pellucida DNA sequences described above were obtained fromovarian cDNA libraries screened with specific zona pellucida antibodiesor known zona pellucida DNA probes. Comparison of isolated sequences topublished protein or DNA sequences and with other clones as they wereisolated was used to classify and identify the clones as describedabove.

The term "zona pellucida protein" is meant to include full lengthproteins ZPA, ZPB, and ZPC, as well as expected variants,immunologically active fragments or peptides contained within theseproteins. The term "zona pellucida DNA" is meant to include thosenucleic acid sequences encoding zona pellucida protein or fragmentsthereof.

The three major classes of mammalian zona pellucida proteins have beendetermined on the basis of homology within the DNAs encoding ZP proteinsof a variety of mammalian species. ZPA includes those peptidespreviously, variously described in the literature as ZP1, ZP2, and ZP4;ZPB includes those peptides previously, variously described as ZP3α andrc 55; and ZPC includes those peptides previously variously described asZP3β and ZP3.

The homology of various species of zona pellucida proteins within aspecific class as compared with a consensus sequence for each class isshown in Table 1. The consensus sequence was derived using theMicrogenie® Sequence Analysis Program (Beckman Instruments, Inc. SpincoDivision, Palo Alto, Calif.). The minimum percent of aligned sequenceswhich must have the same residue at a given position for that residue tobe included in the consensus sequence was 50%. The DNA sequencescorresponding to the amino acid consensus sequences for ZPA, ZPB, andZPC proteins are set out in SEQ ID NOS 25, 26, and 27, respectively.

                  TABLE 1                                                         ______________________________________                                        HOMOLOGY OF DEDUCED ZP PROTEINS AMINO ACIDS                                              ZPA          ZPB     ZPC                                           ______________________________________                                        DOG        78.9%        --      77.3%                                           CAT 78.4% 70.9% 77.5%                                                         COW 77.2% 80.4% 77.2%                                                         PIG 73.0% 77.8% 79.0%                                                         RABBIT 70.1% 74.6% 71.3%                                                      MOUSE 61.6% -- 69.6%                                                          HUMAN -- -- 76.9%                                                             HAMSTER -- -- 70.5%                                                         ______________________________________                                    

The deduced amino acid sequences of the various species of zonapellucida proteins suggest approximate unglycosylated molecular weightsof 75 kD, 55 kD, and 45 kD for ZPA, ZPB, and ZPC, respectively. A moredetailed analysis of both DNA sequence homology and deduced amino acidsequence homology is set out as Examples 13, 14, and 15.

It has surprisingly been found that administration of a specific classof zona pellucida protein to a host animal results in a specificimmunocontraceptive effect and that selection of the appropriate ZPprotein for administration allows induction of desired contraceptiveresults, in terms of permanent sterility or transient infertility. Forexample, vaccination of an animal with zona pellucida protein C inducesantibody titers in that animal which recognize endogenous ZPC resultingin loss of oocytes from the animal's ovary, thereby causing permanentsterility. In contrast, vaccination of an animal with zona pellucidaprotein A, B or combinations thereof induces antibody titers which donot recognize ZPC, but recognize ZPA and/or ZPB. This results incycling, infertile animals for the time period during which anti-ZPAand/or anti-ZPB antibody titers remain high. When such antibody titersfall, the infertility effect is diminished, and the animal regainsfertility.

Vaccination with the purified, isolated, and characterized ZPA, ZPB, orZPC proteins is seen to exert a specific effect on the immunized animalif an autoimmune response is triggered wherein the autoantibodiesgenerated specifically recognize the immunized animals' own specificzona pellucida protein. This self-recognition for antibodies inducedaccording to the present invention may be defined and characterized bythe ability of serum antibodies to recognize at least one epitopepresent on a homologous species zona pellucida protein.

In the preferred method of the invention, an animal is immunized with arecombinant ZPA, ZPB, or ZPC or fragments thereof. The recombinantprotein or peptide may be of homologous species or derived from aheterologous species zona pellucida which shares common epitopicdeterminants, with the proviso that such common epitopic determinantsfunction to induce the desired autoimmune response.

The recombinant protein or peptide fragment may be chemically conjugatedto immune enhancing agents such as Keyhole Limpet Hemocyanin (KLH), andMuramyl dipeptide (MDP), and the like, or alternatively may be providedin the form of a fusion protein, e.g., with foreign protein amino acidsat the amino and/or carboxy terminus. Fully conventional methods forstimulating the production of antibodies upon administration of theproteins or fragments of this invention are well known; similarly,passive immunization techniques involving administration of antibodiesper se, e.g., anti-ZPA antibodies, anti-ZPB antibodies, or anti-ZPCantibodies, to the zona pellucida proteins or fragments of thisinvention is also within the scope of the invention. For details, seeDean, PCT Application WO90/15624 whose disclosure is entirelyincorporated by reference herein.

Thus, to induce permanent sterility in a dog, recombinant canine ZPC maybe employed which is expressed as a bacterial fusion protein (orconjugated to immune enhancing agents) wherein active canine ZPC proteinis conserved and available for interaction with antigen presentingcells. The expressed protein is then administered to a host dog andinduces an autoimmune response in which generated antibodies recognizecanine zona pellucida protein C. This autoimmune effect, whichspecifically recognizes dog ZPC protein or its aggregates, inducespermanent sterility in the vaccinated dog, which sterility is associatedwith a loss of oocytes from the dog's ovary.

Alternately, a non-homologous species ZPC, such as recombinant porcineZPC or peptides thereof which are cross-reactive with canine ZPC, can beadministered to a dog to achieve similar sterilizing effects. Thesterilizing effect, however, is only realized when antibodies capable ofrecognizing the host's own native zona pellucida are induced (oradministered in the context of passive immunization).

In an alternative embodiment of the present invention, theadministration of a host species' own A and/or B class zona pellucidaprotein, or a related A and/or B protein from another species whichinduce antibodies against the host's ZPA and/or ZPB proteins results inan infertility effect which is distinct from that produced by ZPC classantigens. The physiological effect of vaccination with the ZPA and ZPBproteins is a transient one. "Transient infertility" is herein definedas infertility which is maintained when antibodies against self-zonapellucida proteins are sustained in the host animal's circulation at acontraceptively effective concentration (e.g., at titers ofapproximately 1:250 in the dog) and which infertility is diminished whenantibodies against self fall below a contraceptively effective lowerlimit. The reduction in antibodies against self-zona pellucida resultsin restoration of fertility without evidence of major physiologicalchanges in the ovary. Typically, the reduction in antibody titers occurby natural processes in the mammalian host, but other methods ofreducing antibody titers are within the scope of the invention.

Contraceptively effective antibody titers against self zona pellucidaproteins A and B required to maintain infertility will vary with thespecies of vaccinated animal as well as with the species of recombinantZPA or ZPB peptide administered, but may readily be determined, forexample, by testing a panel of the desired animal species with varyingdoses of the specific antigen, measuring the induced titer of anti-selfantibodies by known ELISA techniques, and correlating the titers withreproductive indicators, e.g., cycling, hormone levels, and the like. Ingeneral, antibody titers greater than 1:250 are contraceptivelyeffective.

Based on amino acid sequence homologies, it is expected that all zonapellucida proteins of a particular class contain functional epitopeswhich are cross-reactive between mammalian species. However, absentcharacterization and identification of such functional cross-reactiveepitopes, a preferred, selective contraceptive agent is a homologousspecies zona pellucida protein or antibody thereto.

The present invention will be more completely understood uponconsideration of the following illustrative examples of the practicethereof wherein: Example 1 addresses the isolation of DNAs encodingporcine species ZPA, ZPB and ZPC; Example 2 relates to isolation ofrabbit ZPC DNA; Example 3 relates to isolation of DNAs encoding canineZPA and ZPC; Example 4 addresses isolation of feline DNAs encoding ZPA,ZPB and ZPC; Example 5 relates to cloning and isolation of DNAs encodingbovine species ZPA, ZPB and ZPC; Examples 6 and 7 describeimmunocontraceptive treatment of dogs with naturally-derived porcinezona pellucida proteins; Example 8 relates to serochemical studies onanimals treated in Examples 6 and 7; and Examples 9 and 10 addressrecombinant production of a canine ZPC fusion protein and itsimmunocontraceptive use in dogs. Example 11 relates to the isolation ofDNAs encoding human ZPA and ZPB by methods described herein. Example 12relates to the isolation and sequencing of DNAs encoding cynomolgusmonkey ZPA, ZPB and ZPC. Examples 13-15 relate to the comparison of theDNA sequence and the deduced amino acid sequence of mammalian ZPA, ZPB,and ZPC, respectively. Example 16 relates to the immunization ofcynomolgus monkey using HSPZ and fractionated HZPC. Example 17 relatesto the mapping of mammalian zona pellucida protein epitopes. Example 18describes the immunization of dogs using recombinant ZPC proteins.Example 19 relates to the vaccination of cows and cats with recombinantZP proteins.

Example 1 Isolation of DNA Sequences Encoding Porcine Zona PellucidaProteins ZPA, ZPB, and ZPC

A cDNA library in λgt11 was commercially prepared by Clone Tech, PaloAlto, Calif., from an ovary isolated from a 14 week old pig and wasscreened using an anti-ZP3β antibody obtained from E. C. Yurewicz anddescribed in Keenan et al., Biol. Reprod., 44:150-156 (1991). Eightcandidate clones were identified.

A degenerate DNA oligonucleotide probe (19bps) was constructed torepresent all possible sequences of a short portion of the N-terminusporcine ZP3β as described in Yurewicz et al., J. Biol. Chem.,262:564-571, (1987). The degenerate probe sequence is set out in SEQ IDNO. 28.

Southern analysis of the eight candidate clones isolated by expressionscreening with the degenerate DNA oligonucleotide probe resulted inhybridization with two of the eight candidates. The two clonesrecognized by the degenerate probe were then subcloned into the pBS KSplasmid (STRATAGENE Cloning Systems, La Jolla, Calif.) for sequenceanalysis using the sequence enzyme and the protocol described in theSEQUENASE® Manual (U.S. Biochemical, Cleveland, Ohio). One of theclones, B-8, having an insert size of approximately 1200 base pairs,included a sequence homologous to the N-terminal sequence of mouse ZP3,previously identified by Ringuette et al., Dev. Biol., 127:287-295,(1988). The remaining clone, B-6, had an insert size of approximately1000 base pairs. Neither hybridizing clone contained the C-terminalportion of the gene, as suggested by the lack of homology to the mouseZP3 gene in this region.

The 14-week porcine ovarian library was then rescreened by DNAhybridization. Approximately 150,000 PFUs were plated on agar plateswith E. coli Y1090. After overnight incubation at 37° C., nylon membranelifts of plaques were prepared and screened using the B6 and B8 clonesderived above isolated by screening with the degenerate oligonucleotideprobe set out in SEQ ID NO. 28.

Filters were prehybridized in a solution containing 5× saline, sodiumphosphate, EDTA buffer (SSPE), 5× Denhardt's Reagent, 100 μg/ml salmonsperm DNA, 30% formamide and 0.5% SDS for three hours at 42° C.Approximately 50 ml of the prehybridization solution was used for 12filters (132 mm). After prehybridization, 10 ng of freshly radiolabeledDNA probe in 30% formamide, 5× SSPE was added. The probes were heatdenatured at 95° C. for 3-5 minutes and hybridization with the DNAprobes continued overnight at 42° C. The hybridized filters were thenwashed twice with 100 ml of 5× SSPE at 55° C., for approximately onehour each wash. The filters were then rinsed with 250 ml of 5× SSPE atroom temperature and allowed to air dry. The dried filters were exposedto x-ray film at -70° C. using intensifier screens for at least eighthours and the films were developed for visual analysis.

Among the additional clones isolated were two clones including theC-terminal portion of the porcine ZP3β gene. One clone, λ5-1, wassubcloned into plasmid pBS KS and sequenced. This plasmid, termed pZ57,contained a ZP DNA insert having 1266 base pairs and appeared to encodethe full length amino acid sequence of porcine ZP3β as compared withknown mouse ZP3. Alignment of the deduced amino acid sequence of theclone with the known N-terminal amino acid sequence of ZP3β reported byYurewicz et al., J. Biol. Chem., 262:564-571 (1987), and an internalpeptide sequence of ZP3β corresponding to amino acids 255-274 asprovided by E. C. Yurewicz confirmed the identity of this clone asencoding porcine ZP3β.

The DNA sequence of this clone, termed porcine ZPC, is set out in SEQ IDNO. 5 and its deduced amino acid sequence is set out in SEQ ID NO. 6.

The 14-week porcine ovarian cDNA library was further screened usingrabbit zona pellucida rc 55 cDNA as a probe [described in Schwoebel etal., J. Biol. Chem, 266:7214-7219, (1991)].

One candidate clone of approximately 1700 base pairs, λ2-1, was isolatedand was transferred into the sequencing plasmid pBS KS. The DNA sequenceand deduced amino acid sequence of the porcine DNA insert was determinedusing the method described in the SEQUENASE® manual (US BiochemicalCorporation, Cleveland, Ohio). The sequenced clone contained 1620 basepairs and included a full length copy of the porcine ZP3α gene asconfirmed by alignment of the deduced amino acid sequence with portionsof the known protein sequence of porcine ZP3α provided by E. C. Yurewiczbetween amino acids 206-222, 271-279, and 328-344. The DNA sequence ofthis clone, termed porcine ZPB, is set out in SEQ ID NO. 3. Its deducedamino acid set out in SEQ ID NO. 4.

The 14-week porcine ovarian library was further screened using theprocedure described above and using a DNA probe encoding canine ZPAprotein (as obtained in Example 3 below, SEQ ID NO. 9). A single clone,λ3-5 having approximately 1300 base pairs, was obtained representing theN-terminal 60% of the theoretical porcine ZPA gene as estimated by thesize of the clone in relation to the ZP2 gene isolated from mouse byLiang et al., Mol. Cell. Biol. 10:1507-1515 (1990), and rabbit byDunbar, U.S. Pat. No. 4,996,297, and dog (see Example 3 below).

This clone was then used to rescreen the porcine ovarian library. Threeadditional clones were obtained, two small clones and one clone largeenough to contain the full length sequence. The large candidate clone,λB, having approximately 2200 base pairs, was sequenced, and the datashowed this ZPA clone to lack only approximately seven base pairs of thefull length sequence including the ATG start codon when aligned with themouse ZP2 gene and the canine ZPA gene described in Example 3. The DNAsequence of this clone, termed porcine ZPA, is set out in SEQ ID NO. 1.Its deduced amino acid sequence is set out in SEQ ID NO. 2.

This isolated porcine clone included sequences corresponding topublished sequences of three identified porcine zona pellucida proteins,ZP1 (80 kD), ZP2 (62 kD) as disclosed in U.S. Pat. No. 4,996,297 toDunbar and ZP4 (21 kD) as disclosed by Hasegawa et al., Abst. No. 382,Meeting Soc. Study Reprod. July, 1991. These results suggest that asingular clone encodes one zona pellucida protein which previously hadbeen thought to exist as three separate proteins, i.e., ZP1, ZP2, andZP4. This further suggests that only three major porcine zona pellucidagenes encode three major zona pellucida proteins which here are termedZPA, ZPB, and ZPC. ZPA includes those proteins previously identified asZP1, ZP2, and ZP4. ZPB corresponds to ZP3α and ZPC corresponds topreviously identified ZP3β. Yurewicz et al. J. Biol. Chem., 262:564-571,(1987).

EXAMPLE 2 Isolation and Purification of DNA Sequences Encoding RabbitZPC Protein

Ovaries were removed from five week old rabbits and mRNA was preparedusing the Fast Track™ mRNA isolation kit in accordance with theprocedure described in the Fast Track™ instruction manual, version 3.1,catalog No. K1593-02 (Invitrogen, San Diego, Calif.). A LambdaLibrarian™ kit (Invitrogen, San Diego, Calif.) was used to prepare cDNAand to clone cDNAs into λgt10 according to the manufacturer'sinstructions. Approximately 150,000 PFUs were plated on agar plates withE. coli Y1090. After overnight incubation at 37° C., nylon membranelifts of colonies were prepared and screened with a porcine ZPC DNAprobe using the screening procedures described for Example 1. The probeused was the porcine ZPC sequence as set out in SEQ ID NO. 5.

Two positive clones, λR4 and λR5, hybridized with the porcine ZPC DNA.The size of each of these clones as estimated in agarose gels wasapproximately 1300 base pairs. Both λR4 and λR5 were sequenced asdescribed for Example 1. The sequences were identical except that λR5contained four additional nucleotides at the 5' end. The determined DNAsequence was approximately 75% homologous to the DNA sequence encodingporcine ZPC.

The DNA sequence encoding rabbit ZPC protein is set out in SEQ ID NO. 7.Its deduced amino acid sequence is set out in SEQ ID NO. 8.

Rabbit ZPA and ZPB proteins have been previously identified by Dunbar inU.S. Pat. No. 4,996,297 as P2 and P3, respectively.

EXAMPLE 3 Isolation of DNA Sequences Encoding Canine Zona PellucidaProteins ZPA and ZPC

A 16 week canine ovarian cDNA expression library was commerciallyprepared by Clone Tech, Palo Alto, Calif., in λgt11 generally followingthe methods described in Example 1. The canine ovarian cDNA library wasscreened using antibodies raised against heat solubilized canine zonapellucida. Heat solubilized canine zona pellucida (HSDZ) was preparedgenerally following the procedures described in Dunbar et al.Biochemistry, 19:356-365, (1980) except ganged razor blades were used tomince the ovaries.

Rabbits were immunized with 250 μg HSDZ and 250 μg MDP. Two additionalboosts followed at approximately three week intervals. The resultantrabbit serum was used to screen the canine ovarian cDNA expressionlibrary. Seven candidate clones were obtained. Cross-hybridizationexperiments were performed by Southern blot analysis as follows. Thelargest clone, λ26-1, having approximately 1300 base pairs, was firstused as a probe against all of the other clones in Southern blots. Threeother clones were identified. The largest of the remaining clones, λ20-1and λ7-1, having approximately 800 and 1000 base pairs respectively,were then used as probes in Southern blots. These probes identified noadditional clones. This cross hybridization analysis of the sevencandidate clones to each other indicated that four of these clones wererelated, e.g. four clones hybridized to λ26-1 while the remaining threeλ20-1, λ7-1, and λ19-3 were independent.

The largest of the four related clones, λ26-1, was subcloned into pBS KSplasmid for sequence analysis according to the procedure described inExample 1. The analyzed sequence demonstrated the presence of a longopen reading frame of 1278 base pairs encoding a protein ofapproximately 426 amino acids. Comparison of the deduced amino acidsequence of this clone with the sequences of known zona pellucidaproteins, indicated this clone encoded a protein related to mouse ZP3(ZPC) as reported by Ringuette et al., Dev. Biol. 127:287-295 (1988),hamster ZP3 as reported by Kinloch et al., Dev. Biol., 142:414-421(1990), human ZP3 as reported by Chamberlin et al., Proc. Natl. Acad.Sci. USA 87:6014-6018 (1990) and porcine ZPC protein (see Example 1).The DNA sequence of this clone, termed canine ZPC, is set out in SEQ IDNO. 11. Its deduced amino acid sequence is set out in SEQ ID NO. 12.

The remaining three independent candidate clones were subcloned into thepBS KS plasmid for sequence analysis as described above. The determinedsequence of the 800 base pair clone, 20-1, was compared with known ZPsequences by computer analysis as described above and was found to berelated to the mouse ZP2 (ZPA) [Liang et al., Mol. Cell. Biol.10:1507-1515 (1990)] and porcine ZPA (see Example 1).

The 800 base pair fragment from λ20-1, was then used as a hybridizationprobe to rescreen the canine cDNA library. Two additional candidateclones were identified, the larger of which, λ7A, having approximately2800 base pairs, was subcloned into pBS KS plasmid for sequenceanalysis. Comparison of this sequence with known sequences encoding zonapellucida proteins suggested the candidate clone λ7A contained a fulllength ZPA sequence, but an incorrect N-terminal sequence, e.g., theclone contained an additional 600 base pairs as determined by alignmentwith known mouse ZP2 and rabbit ZPA sequences referenced in Example 1.The second candidate clone, λ9-2, having approximately 1000 base pairs,was then subcloned into the plasmid pBS KS and sequenced. The sequenceof the second clone indicated the presence of a correct N-terminalsequence, but included only approximately the N-terminal 40% of the fulllength clone as determined by alignment with the mouse ZP2 and rabbitZPA genes. Overlap of the two cDNA clones, however, provided the fulllength sequence.

The appropriate pieces of each clone were subcloned as follows togenerate the correct full length zona pellucida clone containing a 2028base pair open reading frame encoding a protein of approximately 676amino acids. The λ7A DNA was digested with Eco RI to yield two insertfragments (2000 bps and 800 bps). These two fragments were eachsubcloned into pBS KS yielding pZ36 and pZ37, respectively. Plasmid pZ37carried the C-terminal portion of this sequence. The λ9-2 DNA insert wasremoved from the λ vector and subcloned into pBS KS to yield pZ38.Plasmid pZ36 was digested with Hind III to remove approximately 1350 bpsof the N-terminal portion of the λ7A gene fragment (about 850 bps ofnonsense DNA and 500 bps of coding sequence). This digestion alsoremoved one of the Eco RI insert ends and left a single Eco RI site. ThepZ37 Eco RI insert was then moved into the single remaining Eco RI sitein the modified pZ36 (pZ36 Δ1) to reestablish the relative DNA structureorientation that existed in the λ7A insert (1450/2800 bps). Thiscombined plasmid was then opened with Hind III and the Hind III fragmentfrom pZ38 carrying the N-terminal ZP DNA sequence was inserted to createplasmid pZ39 which is a pBS KS carrying the full length canine ZPAsequence. The DNA sequence of this canine ZPA gene is set out in SEQ IDNO. 9. Its deduced amino acid sequence set out in SEQ ID NO. 10.

EXAMPLE 4 Isolation of DNA Sequences Encoding Feline Zona PellucidaProteins ZPA, ZPB, and ZPC

Ovaries were isolated from five cats approximately three to four monthsin age. Messenger RNA was isolated from six ovaries using the FastTrack™ mRNA Isolation Kit (Invitrogen, San Diego, Calif., Catalog No.K1593-02) using the protocol provided with the kit. cDNA was preparedusing the protocol and cloned into λgt10 as described in Example 2.

Approximately 150,000 plaque forming units (PFUs) were plated on agarplates with E. coli Y 1090. After overnight incubation at 37° C., nylontransfer membranes were used to prepare and screen plaque lifts. Plaqueswere screened using a mixture of DNA probes in equal proportionsencoding porcine ZPA, ZPB, and ZPC proteins and using the hybridizationprocedure as described for Example 2. A total of 81 positive clones wereidentified. Twelve of these clones were plaque-purified. Southernanalysis of these clones using porcine ZPA, ZPB, and ZPC DNAsindividually as probes indicated that seven of these clones encoded ZPCproteins and one clone encoded a ZPA protein. Four of the clonescontained inserts which could not be separated by Eco RI digestion.

Five of the ZPC clones were between 1200-1350 base pairs in length. Oneclone, λC-112, having approximately 1350 base pairs was subjected tosequence analysis as described above and its deduced amino acid sequencewas found to be approximately 70% homologous to the canine ZPC proteinobtained in Example 3. The DNA sequence of this feline ZPC clone is setout in SEQ ID NO. 17. Its deduced amino acid sequence is set out in SEQID NO. 18.

The single feline ZPA clone, λC-116, was sequenced and found to beapproximately 2215 base pairs in length. The deduced amino acid sequencewas approximately 75% homologous to the canine ZPA protein characterizedin Example 5. The DNA sequence of this feline ZPA clone is set out inSEQ ID NO. 13. Its deduced amino acid sequence is set out in SEQ ID NO.14.

The remaining 69 positive clones were rescreened using porcine ZPB DNAas a probe (SEQ ID NO. 3). Ten positive clones were obtained. Thelargest clone, λC-1, contained approximately 1.7 kilobases as determinedby agarose gel electrophoresis. This clone was sequenced, and itsdeduced amino acid sequence was found to be approximately 80% homologousto the porcine ZPB protein described in Example 1. The DNA sequence ofthis feline ZPB clone is set out in SEQ ID NO. 15. Its deduced aminoacid sequence is set out in SEQ ID NO. 16.

EXAMPLE 5 Isolation of DNA Sequences Encoding Bovine ZonaPellucida-Proteins ZPA, ZPB, and ZPC

A cDNA library was constructed from a five month bovine ovary by themethod described in Example 2. The bovine ovarian library was screenedwith DNA hybridization probes representing each of the classes of zonapellucida proteins using a mixture of equal proportions of porcine DNAprobes encoding ZPA (SEQ ID NO. 1), ZPB (SEQ ID NO. 3), and ZPC (SEQ IDNO. 5) proteins, as described for Example 2 and using the proceduresdescribed for Example 1. Initial screening yielded three candidateclones. Southern analysis of these clones with individual porcine ZPA,ZPB, and ZPC DNA probes used in the initial screening indicated that oneof the clones, λB2, having approximately 650 base pairs, encoded ZPA. Asecond clone, λB-1 having approximately 1000 base pairs encoded ZPB. Athird clone, λB14, having approximately 1200 base pairs, encoded ZPC.

The bovine ovarian library was then rescreened with the mixed porcine ZPDNA probes. Two additional clones were obtained and identified bySouthern analysis as encoding ZPC.

The Eco RI inserts of the ZPA, ZPB, and largest ZPC clone were subclonedand their DNA sequences analyzed. The sequences encoding these bovineZPA, ZPB and ZPC fragments were set out in SEQ ID NOS. 19, 21, and 23,respectively. Their deduced amino acid sequences are set out in SEQ IDNOS. 20, 22, and 24, respectively.

EXAMPLE 6 Immunization of Dogs with Heat-Solubilized FractionatedPorcine Zona Pellucida

Heat-solubilized, porcine zona pellucida (HSPZ) was prepared generallyfollowing the procedures described by Dunbar et al. Biochemistry,19:356-365, (1980) but using a hand powered meat grinder instead of theZonamatic described. Following isolation, the zona pellucida protein wassolubilized in 0.1 M sodium carbonate buffer, pH 9.6, and was dialyzedextensively against 6M urea. The resultant solution, a volume of 2-3 mlcontaining approximately 12 μg of HSPZ, was subjected toisoelectric-focusing in a BIORAD Rotofor isoelectric-focusing chamber asfollows. An isoelectric gradient was established using 1% ampholyteshaving a pI range of 3-10. The zona pellucida protein was introducedinto the mid-range chamber (pI 7.0) and allowed to focus forapproximately four hours at 4° C. or until the voltage stabilized.

Twenty isoelectrically focused fractions were collected and analyzed bySDS PAGE and Western blot analysis for pig zona pellucida proteins.Acidic fractions having a pI range of approximately 3.5-5.5 and whichcontained the porcine zona pellucida proteins were combined. Thefractions were dialyzed into 0. 1M carbonate buffer, pH 9.6 andconcentrated to approximately 3 mg/ml. This antigenic preparation wasused to vaccinate animals as described below. Analysis of this antigenicpreparation by two-dimensional gel electrophoresis indicated thepresence of ZPA and ZPB protein. However, ZPC was not revealed to bepresent in this preparation.

The HSPZ antigenic preparation was added to a 50/50 water oil emulsionwith incomplete Freund's adjuvant (Sigma, St. Louis, Mo.) containing 250μg of MDP per dose. One ml of the 50/50 water oil emulsion contained0.425 ml paraffin oil, 0.075 ml mannide monooleate, and 0.5 ml PBScontaining 250 μg threonyl-MDP (SYNTEX Corporation) and the amount ofHSPZ described in Table 3 below.

Four random breed dogs aged 10-12 weeks were immunized with HSPZ usingthe regimen described in Table 2.

                  TABLE 2                                                         ______________________________________                                                          mg HSPZ                                                     ______________________________________                                        Prime          Time 0   0.1                                                     Boost #1 Week 4 1.0                                                           Boost #2 Week 8 0.25                                                          Boost #3 Week 12 0.2                                                          Boost #4 Week 16 1.0                                                          Boost #5 Week 36 1.0                                                        ______________________________________                                    

The antisera produced by these animals was monitored via ELISAmethodology. By week 17 antibody titers against self, e.g. againstcanine zona pellucida proteins, had reached a maximum (8-16K by ELISA)and thereafter began to drop.

At week 36, one animal was unilaterally ovariectomized and the removedovary was sectioned and stained with periodic acid schiff stain (PAS)for histological examination. The ovary appeared normal, as evidenced bythe presence of follicles in all stages of development. At week 52, twoof the four test animals were observed to exhibit estrus behavior. Theremaining two test animals exhibited estrus behavior at approximatelyone and a half years when the first two test animals experienced theirsecond heat. All test animals were bred repeatedly with competent malesand by artificial insemination, however, none became pregnant. Duringthis same period, animals in various test regimens in which no selftiters were obtained, as described in Example 10, became pregnant whenpresented with the same males or artificial insemination techniques.

Two weeks following the breeding sessions, e.g. at 54 weeks, the twoearly cycling animals were unilaterally ovariectomized and the removedovaries were sectioned for histological examination. The ovariesappeared normal for this stage of follicular activity despite thefunctional infertility demonstrated.

EXAMPLE 7 Vaccination With Porcine ZPC Protein

A purified porcine ZPC protein (ZP3β) was obtained from E. Yurewicz,prepared as described in J. Biol. Chem., 262:564-571, (1987).

Vaccines were prepared by adding 167 μg purified porcine ZPC protein(ZP3β) to a 50/50 water-oil emulsion with complete Freund's adjuvant(Sigma No. F5881, St. Louis Mo.), for the priming dose or withIncomplete Freund's Adjuvant (Sigma No. F5506, St. Louis, Mo.)containing MDP as described in Example 6 for the booster doses.

Five random breed dogs of approximately 10-12 weeks of age were injectedwith the ZPC vaccine preparation described above using the regimendescribed in Table 3.

                  TABLE 3                                                         ______________________________________                                                        mg of ZPC                                                     ______________________________________                                        Prime        Time 0   0.167                                                     Boost Week 3 0.167                                                            Boost Week 6 0.167                                                            Boost Week 28 0.167                                                         ______________________________________                                    

Each animal's antibody titer versus self- zona proteins, e.g., versuscanine zona pellucida proteins, was monitored by ELISA, using the methoddescribed in Dunbar, Two Dimensional Gel Electrophoresis andImmunological Techniques, 1987. ELISA microtiter plates were coated withHSDZ in antigen-coating buffer (0.1M sodium carbonate, pH 9.6).Biotinylated rabbit-antidog IgG was used as the second antibody. ABCreagent (Avidin-biotinylated peroxidase complex) and O-phenylene diaminedihydrochloride with a peroxide substrate was used for visualization.Only two animals produced antibodies versus self achieving peakself-antibody titers of 16K by week 4. The other three animals producedno self-antibody titers but achieved peak antibody titers of 4K againstporcine zona pellucida protein. During the period of time between week20 and week 36, all dogs were observed to exhibit estrous behavior. Theanimals were bred repeatedly with proven males. Only the two animalshaving antibody titers versus self zona pellucida proteins remainedinfertile. All other animals in the study became pregnant.

Two weeks after estrous and breeding the two infertile dogs exhibitingself-antibody titers were unilaterally ovariectomized and the removedovaries were sectioned and stained with PAS for histologicalexamination. The histological examination revealed abnormal morphologyin the ovaries of the infertile dogs. No evidence of ongoingfolliculogenesis was seen and the ovaries were depleted ofoocyte-containing follicles. In addition, no primordial oocytes wereseen.

EXAMPLE 8 Western Analysis of Antisera Produced by Vaccinated Animals

In an attempt to better understand the immune response and differentphysiological effects obtained in the two studies described in Examples6 and 7, antisera produced in each test group was analyzed by WesternAnalysis against a variety of antigens including natural porcine ZPC,heat-solubilized dog zona pellucida (HSDZ), recombinant dog ZPA and ZPC,and recombinant pig ZPC. Western blots were probed with antiserumobtained from the test animals of Example 6, e.g., animals immunizedwith isoelectric focused, heat-solubilized porcine zona pellucida, andwith antiserum obtained from the two test animals of Example 7 whichcontained antibodies against self-zona proteins.

The data demonstrate no recognition of recombinant porcine or canine ZPCby antisera from infertile, but cycling dogs immunized with heatsolubilized porcine zona pellucida which contained no demonstrable ZPCby PAGE analysis, however, natural ZPC, HSDZ and recombinant canine ZPAwere recognized. In contrast, antisera obtained from infertile dogswhose ovaries were depleted of oocytes recognized recombinant ZPCprotein, i.e., the polypeptide backbone.

A key difference in the antibody recognition of antigen was that onlythe antisera obtained from dogs having ovaries devoid of oocytesappeared to recognize the recombinant dog ZPC antigen. Infertile dogswhose antisera strongly recognized natural ZPC, HSDZ, and recombinantdog ZPA demonstrated no recognition of recombinant dog ZPC.

Given that autoimmunity is essential for a contraceptive effect, thesedata suggest that infertility without histologically evident ovariandysfunction can be obtained in dogs via an autoimmune response againstdog ZPA antigens. In contrast, histologically confirmed ovariandysfunction, i.e., loss of oocytes, which would result in permanentsterility, requires the generation of antibodies which specificallyrecognize homologous species ZPC protein.

EXAMPLE 9 Expression of Recombinant ZP Proteins

I. Construction of Expression Vectors

The plasmid vector pZ90 shown in FIG. 1 was constructed from fragmentsof the plasmids pUC9 (Vierra & Messing, Gene 19:259-268 (1982)) andpβga12 (Queen, J. Mol. App. Gen. 2:1-10 (1983)). The single Pvu IIrestriction site present in pβga12 was converted to a Sal I site using aSal I polylinker adaptor purchased from New England Biolabs. The DNAsequences between the new Sal I site and a pre-existing Sal I site wereexcised by digestion with Sal I, religated and screened for the reducedsize plasmid.

A Cla 1-Nde I fragment of the modified pβga12 plasmid which carried theλCI repressor gene, the λpR promoter and the Lac Z gene(β-galactosidase) was inserted into pUC9 between its Acc I and Nde Irestriction sites. The pUC9 plasmid carries the ampicillin resistance(Amp^(R)) gene and col EI replication origin (ori) needed to maintainthe plasmid in E. coli cells. The combination plasmid was furthermodified to convert the Bam HI site 3' of the ATG initiation codon (ATGGAT CCN) to a Bgl II site 5' of the ATG initiation codon (AGATCTATG).This was accomplished by partially digesting the plasmid with Rsa I. Oneof the several digestion points was about 20 bps 5' of the Bam HIrestriction site. When the partially digested plasmid was digested withBam HI, some of the plasmids produced were nearly full length. Asynthetic oligomer (GTACTAAGGAAGATCTATGGATCC) (SEQ ID NO. 29) wasproduced to replace the sequence that had been removed(GTACTAAGGAGGTRGTATGGATCC) (SEQ ID NO. 30). The net effect of thisreplacement was the substitution of 3 bps to create the Bgl IIrestriction site. A DNA fragment containing approximately 3000 basepairs of the Lac Z gene was then excised by restriction digestion withBgl I and Ban II and was followed by insertion of a synthetic oligomercontaining a Bam HI site. The plasmid was cut with Bgl I and Ban II, andthen treated with nuclease S1 to create blunt ends. A Bam HI linker (NewEngland Biolabs) was inserted at the blunt ends of the digested plasmid.Next a Pvu II restriction site between the λCI repressor gene and theori sequence was converted to a Hind III site using a synthetic linker.The Pvu II restriction site was cut with Pvu II, and a Hind III linker(New England Biolabs) was ligated to the blunted ends. Because theremaining lac Z sequence was missing the first 8 codons of the naturalsequence, these 8 codons were replaced by synthesizing a syntheticoligomer that began with a Bgl II site and encoded the lac Z wild typegene product (βgal) N-terminal sequence.

The synthetic oligomer was prepared by synthesizing four oligomershaving the sequences set out in SEQ ID NO. 31 (oligomer 1), SEQ ID NO.32 (oligomer 2), SEQ ID NO. 33 (oligomer 3), and SEQ ID NO. 34 (Oligomer4). Oligomers 2 and 3 were phosphorylated by treating with-kinase andATP to add phosphate to the 5' end. Oligomers 1 and 2 were thenhybridized to oligomers 3 and 4, respectively, by incubation at 100° C.followed by a slow cooling in 200 μM NaCl. The resultant oligomer hadthe sequence set out in SEQ ID NO. 35. The synthetic oligomer as set outin SEQ ID NO. 35 had Bgl II-Pvu II ends and was substituted for the BglII-Pvu II sequence of the plasmid by restriction digestion of theplasmid and ligation with the oligomer.

The resultant plasmid was termed pZ90 and is shown in FIG. 1. Theplasmid pZ90 can be used to express recombinant proteins by heatinduction, using the heat labile λCI repressor. The heat-induciblerepressor and promoter of pZ90 was next replaced with the chemicallyinducible promoter ptac (Amann et al., Gene 25:167-178 (1983)). The ptacpromoter is controlled by the lac repressor, a product of the lac I gene(Farabaugh, Nature 279:765-769 (1978)). The Lac I gene was obtained frompMC9 (Miller et al., The EMBO Journal 3:3117-3121 (1984)) by use of PCRmethodology as described by Innis and Gelfand, In: PCR Protocols: AGuide to Methods and Applications, Innis, M. A., Gelfand, D. H.,Sninsky, J. J. and White, T. J. (eds)., pgs 1-12, Academic Press, Inc.,San Diego, Calif. The primers used were complimentary to the Lac Ipromoter at one end and the Lac I gene termination codon at the oppositeend. The N-terminal primer carried a Hind III site and the C-terminalprimer carried a tac promoter sequence followed by a Bgl II site. TheN-terminal primer had the sequence set out in SEQ ID NO. 36. TheC-terminal primer had the sequence as set out in SEQ ID NO. 37 whichincludes a Dra 3 site having the sequence 5'-CACAATGTG-3'. The resultinglac I - ptac DNA fragment having Hind III and Bgl II restriction sitesat its respective ends was then used to replace the Hind III--Bgl IIfragment of pZ90 which carried the λCI repressor and pR promotor. Thisreplacement yielded the plasmid pZ98 shown in FIG. 2.

II. Insertion of Recombinant ZP DNA

DNA sequences encoding porcine ZPC were prepared by the PCR proceduresdescribed above (Innis & Gelfand) from the plasmid pZ57 prepared inExample 1, which contains the full length porcine ZPC sequence obtainedfrom λgt11 clone 5-1 described for Example 1. During the PCR procedurethe porcine ZPC gene was modified by using primers that did not includethe leader sequence and the hydrophobic tail. The N-terminal primer usedhad the sequence set out in SEQ ID NO. 38 which included an internal BamHI restriction site having the sequence 5 '-GGATCC-3'. The C-terminalprimer used had the sequence as set in SEQ ID NO. 39 includes a Sal Irestriction site having the sequence 5'-CTCGAG-3' and an internal Xho Irestriction site having the sequence 5'-CTCGAG-3'. The modified ZPC genecontained base pairs 105 to 1154 encoding ZPC amino acids 1-350.

To the 5' end of the modified porcine ZPC gene was added a Bam HIrestriction site, and to the 3' end was added an Xho I site, aHexa-CAT-codon sequence (CAT)₆, a termination codon, and a Sal Irestriction site. This modified porcine ZPC gene was inserted into theBam HI- Sal I restriction site of pZ98 to yield the porcine ZPCexpression vector, plasmid pZ156 shown in FIG. 3. The (CAT)₆ sequenceproduces a C-terminal hexahistidine (His₆) amino acid sequence in therecombinant fusion protein which permits purification of the fusionprotein by immobilized metal in affinity chromatography.

In a similar manner as described above, the plasmid pZ156 when digestedwith Bam HI and Xho I, may be used to receive any other recombinant ZPgene or gene fragment for expression as a βgal fusion protein which canbe purified by metal ion affinity chromatography.

III. Expression of Porcine ZPC Fusion Protein in E. coli

The expression vector pZ156 (FIG. 3) was transformed into E. coli strainTop 10F' (Invitrogen. San Diego, Calif.) by the procedure of Chung etal., Proc. Natl. Acad. Sci. USA 86: 2172-2175 (1989). The transformed E.coli cell line was termed Strain ZI 156, and was used to expressrecombinant porcine ZPC-βgal fusion protein.

Bacterial cultures of ZI 156 were grown in Luria Broth (LB) containing100 μg/ml ampicillin at 30° C. until the cell density reached an OD⁶⁰⁰of approximately 1.5. Isopropyl beta-D-thiogalactopyranoside (IPTG) (3ml of 100 mM solution/1 media) was added to induce expression from thetac promoter, and the cells were further incubated at 30° C. for 2-3hours. The cells were harvested by centrifugation, and the resultingcell pellet was frozen at -70° C.

The frozen cell pellets were suspended in 10 mM EDTA (1 g/2-2.5 ml) andtwice sonicated at 50% power for 3 minutes, cooling in an ice bathbetween each sonication. The cell lysate was then centrifuged at 3300×gfor one hour and the hard pellet was retained. This lysis procedure wasrepeated using the hard pellets.

In order to remove residual EDTA, the final hard cellular pellet wasdispersed in a small volume of water by a brief burst of sonication, thesuspension was centrifuged, and the supernatant discarded. The washedpellet was thoroughly resuspended in Buffer A, (6M guanidinehydrochloride (GuHCl), 100 mM Na H₂ PO₄, 10 mM TRIS pH 8, atapproximately 0.5 ml per original gram of cell pellet). The suspensionwas centrifuged at 10,000×g for 45 seconds and the supernatant wasretained while the pellet was discarded.

The retained supernatant was loaded onto a Ni column (in Buffer A) andthe column was washed with 10 column volumes of Buffer A. The column wasnext washed with 5 volumes each Buffers B-D, each containing 8M urea,100 mM NaH₂ PO₄, and 10 mM TRIS, and having successively reduced pHvalues of 8, 6.3, 5.9 for Buffers B, C, and D, respectively. Therecombinant pZPC-βgal fusion protein eluted with Buffer E, at pH 4.5 asshown by screening by Western Blot analysis using rabbit anti-HSDZ andanti-HSPZ as probes. Further elution may be accomplished using Buffer F(pH 2.5) (8M GuHCl₂ 200 mM Acetic Acid).

The fusion protein obtained by this protocol was prepared in its finaldose for injection into a host animal by adjusting the final volume to0.5 ml in 8M urea, and adding it to 0.5 ml adjuvant as described above.Each dose was injected subcutaneously into a test animal.

EXAMPLE 10 Vaccination of Dogs with Recombinant ZPC-β gal Fusion Protein

Eleven mixed breed dogs approximately 5-6 months of age were randomlyselected from test animals previously treated at approximately 2 monthsof age with heat solubilized porcine zona pellucida orchromatographically purified porcine ZP3β in combination with variousbiopolymers as adjuvants and drug releasing vehicles. Six weeks postfirst injection, i.e., three and a half months of age, all test animalshad achieved antibody titers versus HSPZ in the range of 2-16K asdetermined by ELISA. However, none of the test animals achieved antibodytiters against self-antigen, e.g., HSDZ.

At 5-6 months of age, five of the test animals were then injected with aloading dose of the porcine ZPC-β gal fusion protein prepared asdescribed for Example 9. The recombinant ZPC-β gal fusion proteinproduced in Example 9 was adjusted to the desired dose in a final volumeof 0.5 ml 8M urea and combined with 0.5 ml adjuvant. The adjuvant,N-acetyl-D-glucosaminyl-β(1,4)-N-acetyl muramyl-L-alanyl-D-isoglutamine(GMDP), 250 μg, was dispersed in 0.42 ml mineral oil, 0.157 ml L-121block polymers, and 0.02 ml Tween 80. Each dose was injectedsubcutaneously into the five test animals. The remaining 6 animals weremaintained as controls.

Following a total of four injections given at 2-3 week intervals,antibody titers versus self antigen, e.g., HSDZ, were obtained in alltest animals, with peaks in the range of 2-8 K as measured by ELISA.

Some of the control animals began to cycle beginning at approximately 9months of age, and by 11 months of age, 4 of 6 control animals hadexperienced their first estrus. In contrast, none of the 5 test animalswhich had received recombinant ZPC-β gal fusion protein had cycledduring this same time period. However, although the first estrus wasdelayed for several months in the test animals, they eventually began tocycle. Two of the five vaccinated dogs became pregnant during theirsecond estrus after immunization while a third dog became pregnantduring its third estrus after immunization; however, the two remainingtest animals remain infertile through three estrus cycles and nearly twoyears after vaccination.

EXAMPLE 11 Isolation of Human DNA Sequences Encoding Human ZonaPellucida Proteins ZPA and ZPB

A human genomic DNA library purchased from Stratagene (catalog no.946203) was used for the isolation of DNA sequences encoding human ZPproteins. The library consisted of 9-23 kb inserts of human DNA (fromplacenta tissue of a male Caucasian) cloned into the Lambda Fix™IIvector (Stratagene). Approximately 40,000 pfus were plated on E. colistrain LE 392 (Stratagene, catalog no. 200266), as described in theStratagene protocol, but replacing MgSO₄ with MgCl₂. After overnightincubation, nylon membrane lifts of the plaques were prepared andscreened with ³² P-labelled porcine ZPA cDNA (SEQ ID NO. 1) and with ³²P-labelled porcine ZPB cDNA (SEQ ID NO. 3) as described in Example 2.

Three clones 1-1, 2-2, and 4-9 were shown to hybridize to the porcineZPB cDNA (SEQ ID NO. 3). Clones 1-1 and 4-9 were deposited with theAmerican Type Culture Collection, (ATCC) 12301 Parklawn Drive,Rockville, Md., on Jan. 27, 1993 under ATCC Accession Nos. 75406 and75405, respectively. Human DNA inserts were isolated from these clonesand analyzed by restriction endonuclease digestion with Eco RI andSouthern blot analysis as described in Example 1. Table 4 shows theresults of Eco RI digestion of these clones.

                  TABLE 4                                                         ______________________________________                                        HUMAN GENOMIC ZPB EcoRI INSERTS                                                    Fragment 1-1          2-2   4-9                                          ______________________________________                                        A                      2.8 kb  2.8 kb                                           B 2.2 kb                                                                      C 2.0 kb                                                                      D 1.5 kb  1.5 kb                                                              E 0.2 kb  0.2 kb                                                              p 3.2 kb 3.2 kb 3.2 kb                                                        G 0.7 kb                                                                    ______________________________________                                    

Southern blot analysis revealed four Eco RI fragments which were judgedto carry ZPB coding sequences based on hybridization to the porcine ZPBcDNA (SEQ ID NO. 3). Clone 1-1 DNA included a 2.2 kb, 2.0 kb, and 1.5 kbEco RI fragments which so hybridized. Clone 2-2 DNA included a 2.8 kbEco RI hybridizing fragment. Clone 4-9 DNA included a 2.8 kb and a 1.5kb Eco RI fragment which hybridized to the porcine ZPB cDNA probe. Allinserts additionally included a 3.2 kb non-hybridizing Eco RI fragment;inserts from clones 1-1 and 4-9 both provided 0.2 kb non-hybridizingfragments; and clone 1-1 additionally provided a 0.7 kb non-hybridizingfragment.

Further restriction analysis revealed the fragment alignment shown inFIG. 4. Six of the fragments (A-F) were subcloned into PBSKS forsequence analysis, as described in Example 1. Preliminary sequenceanalysis confirmed the fragment alignment shown in FIG. 4, and suggestedthat the complete coding sequence of the human ZPB gene may be fromclones 1-1 and 4-9. This was confirmed by nucleotide sequence analysisof the inserts, and comparison of the sequences with the feline ZPBsequence (SEQ ID NO. 15) and porcine ZPB sequence (SEQ ID NO. 3). TheDNA sequence and deduced amino acid sequences for human ZPB are set outas SEQ ID NO. 40 and 41, respectively.

Clones hybridizing to the porcine ZPA cDNA (SEQ ID NO. 1) under theconditions described in Example 1 were also isolated. Two positiveclones, A1 and A4 were identified. The clones were deposited with theAmerican Type Culture Collection, 12301 Parklawn Drive, Rockville, Md.20852, on Jan. 27, 1993 under ATCC Accession Nos. 75404 and 75403respectively. Southern blot analysis revealed that these clones containall or part of the human ZPA gene. DNA was isolated from these clonesand was analyzed by Bgl II, Hind III, and Not I restriction endonucleasedigestion and Southern blot analysis as described in Example 1. The sizeof the A1 clone DNA insert is approximately 11.6 kb, and that of the A4clone is approximately 13.2 kb. Two of the Bgl II fragments whichhybridized with the porcine ZPA cDNA (SEQ ID NO 1) were subcloned intopBSKS for sequence analysis, as described in Example 1. Sequenceanalysis revealed that A1 and A4 collectively contain the human ZPA geneas supported by comparison to sequences with the porcine ZPA cDNA (SEQID NO. 1) and the canine ZPA cDNA (SEQ ID NO. 11). The complete DNAsequence and the deduced amino acid sequence are set out as SEQ ID NOS.42 and 43, respectively.

EXAMPLE 12 Isolation and Sequencing of DNA Encoding Cynomolgus MonkeyZPA, ZPB, and ZPC

Cynomolgus monkey cDNA libraries were constructed in λgt10 as describedbelow. Briefly, a set of ovaries were collected from two femalecynomolgus monkeys aged 1.5 years and 2 years, and a second set fromthree females aged 3 years, 4 years, and 14 years of age. Messenger RNAwas isolated using the Fast Track™ mRNA isolation kit following themanufacturer's instructions. The cDNA was prepared using the LambdaLibrarian™ (Invitrogen, as described in Example 2) kit following theprotocol provided with the kit. The cDNA was packaged into lambda phageheads using the Protoclone® (Promega, Madison, Wis.) λgt10 EcoRI armsplus the Packagene® (Promega) lambda DNA packaging system following themanufacturer's instructions. This procedure generally produced librarieswith a titer of greater than 1×10⁶ pfu/ml. The monkey cDNA library wasthen screened using porcine ZPA, ZPB, and ZPC probes isolated from theporcine cDNA as described in Example 1. Screening was accomplished bypreparing duplicate plaque lifts using Nytran® nylon filters (0.2 μMpore size). The filters were prehybridized in a solution of 5× SSPE(43.83 g/l of NaCl, 6.9 g/l of NaH₂ PO₄, H₂ O, 1.85 g/l of EDTA, pH7.4), 5× Denhardts Reagent (1 g/l of Ficoll [type 400], 1 g/l ofpolyvinylpyrrolidone and 1 g/l bovine serum albumin), 100 μg/mlsonicated, denatured salmon sperm testes DNA, 30% formamide, and 0.5%SDS, for 3 hrs. at 42° C. Radio-labelled probes were prepared using[α-³² p] -dATP and the Prime-a-Gene® (Promega) labelling system. Afterprehybridization, 10 ng of freshly radio-labelled probe was heatdenatured at 95° C. for 5 minutes in 50% formamide and 100 μg/mlsonicated, denatured salmon testes DNA, and was added to the filters.The hybridization was carried out at 42° C. for 15-24 hours. Thehybridized filters were then washed twice with 100 ml of 5× SSPE at 55°C., for approximately one hour each wash. The filters were then rinsedin 250 ml of 5× SSPE at 55° C. and allowed to air dry. The dried filterswere exposed to x-ray film (Kodak XAR5, Eastman Kodak, Rochester N.Y.)at -70° C. using two intensifying screens (Kodak X-OMATIC™) for at leasteight hours. The film was then developed for visual analysis.

Exhaustive screening of the two cynomolgus monkey ovarian cDNA librariesusing all of the porcine probes yielded a total of 12 candidate clones.Southern hybridization revealed that only one of these clones (λCM 4-2)hybridized to the porcine ZPA probe. This clone contained an insert of560 bp. Sequencing of the insert was performed using the Sequenase®Version 2 kit (U.S. Biochemicals, Cleveland, Ohio) according to themanufacturer's instructions. Sequencing revealed that the 560 bp insertwas homologous to the 3' end of other mammalian ZPA genes. The 560 bpfragment represents just under 25% bp of the full-length sequence andcontains an open reading frame of 492 bp which would encode a protein of164 amino acids. The DNA sequence and the deduced amino acid sequence ofthe cynomolgus monkey ZPA cDNA is set out as SEQ ID NOS. 44 and 45,respectively.

Exhaustive screening of the cynomolgus monkey ovarian cDNA librarieswith the porcine ZPB probe yielded a single ZPB candidate clone havingan insert of 866 bp. Sequence analysis suggests that the insert includesthe C-terminal 50% of the expected full-length sequence. The DNAsequence and deduced amino acid sequence of the monkey ZPB insert areset out as SEQ ID NOS. 46 and 47, respectively. Screening of monkeyovarian cDNA libraries with the porcine ZPC DNA probe yielded onlypartial ZPC clones, the largest (λCM1-1) having an insert ofapproximately 1300 bp which contains just over 50% of the C-terminalportion of the full-length sequence based on comparison to known ZPCclones, (particularly the human ZPC clone). The clone contains an openreading frame of 672 bp which would encode a protein of 224 amino acids.The clone also contains stop codons immediately 5' to the codingsequence in all three reading frames. The DNA sequence and the deducedamino acid sequence of the cynomolgus monkey ZPC clones are set out assequence ID NOS 48 and 49 respectively.

EXAMPLE 13 Comparison of ZPA DNA and Deduced Amino Acid Sequences

Table 5 shows a comparison of the DNA and deduced amino acid sequence ofmammalian ZPAs.

                                      TABLE 5                                     __________________________________________________________________________    ZPA HOMOLOGY                                                                  PROTEIN HOMOLOGY                                                                  Mouse                                                                             Rabbit                                                                             Pig Cow Dog  Cat Monkey                                                                            Human                                       __________________________________________________________________________    Mouse                                                                             --  61.0%                                                                              54.2%                                                                             60.8%                                                                             57.9%                                                                              56.9%                                                                             57.2%                                                                             58.9%                                         Rabbit 73.0% -- 63.0% 69.8% 66.2% 64.6% 65.1% 68.9%                           Pig 69.0% 75.6% -- 79.9% 69.6% 70.2% 56.9% 63.9%                              Cow 70.5% 79.0% 86.2% -- 78.3% 77.8% 59.0% 63.6%                              Dog 70.4% 77.2% 80.4% 84.8% -- 83.1% 66.9% 67.5%                              Cat 69.6% 77.5% 81.3% 84.7% 88.9% -- 65.5% 67.4%                              Monkey 56.7% 59.6% 56.6% 57.0% 59.2% 58.4% -- 95.8%                           Human 68.4% 74.6% 73.7% 63.1% 74.4% 75.3% 96.3% --                          __________________________________________________________________________    DNA HOMOLOGY                                                              

Data is presented as a cross-wise comparison of the ZPA protein and DNAsequences. The comparison of the protein sequences are shown in theupper right hand side of the table, above the diagonal dashed lines. Thecomparison of the DNA sequences are shown in the lower left hand side ofthe table, below the diagonal dashed lines. The ZPA DNA and deducedamino acid sequences are highly homologous between species. The homologyis highest between members of the same order within the class mammalia.For example, the human and cynomolgus monkey (primata), the pig and cow(ungulata), and the cat and dog (carnivora) sequences have the mostsimilarity. The high degree of homology between the ZPA genes, as wellas between the ZPB (see Example 14) and ZPC (Example 15) genes from avariety of mammalian species, implies a great deal of structuralsimilarity in the ZP layers of these species. However,post-translational modification differences such as glycosylation andothers, could represent a potential source of variation.

One protein processing site that all of these ZPA proteins have incommon is a furin cleavage site (R-X-R/K-R; Hosaka et al. J. Biol. Chem,266:12127 (1991)) near the C-terminal end of the protein. In fact, withonly a few exceptions, all ZP proteins contain a furin processing sitenear the C-terminus This furin site could serve to cleave off a putativemembrane anchor sequence which would allow the processed proteins tomove toward the outer edge of the growing ZP layer.

The human ZPA gene contains an exon near the 3' end that is present inthe cynomolgus monkey ZPA sequence, but not present in the ZPA genesfrom other species. This extra exon codes for an amino acid sequencethat occurs after the furin processing site, which suggests that theC-terminal fragment generated by furin cleavage might still be importantto the function of the ZP layer or to the oocyte in some way.

There are 20 conserved cysteine residues and one or two non-conservedcysteine residues in each of the full-length ZPA sequences. Thenon-conserved cysteine residues occur either in the N-terminal leadersequence region, or in the extreme C-terminal region of the sequence,where a large amount of the variation between the ZPA sequences occurs.The high degree of homology and the large number of conserved cysteineresidues suggests that the tertiary structures of the ZPA proteins aresimilar.

It has been noted previously that there are regions of homology betweenthe ZPA and ZPB class proteins (Schwoebel et al. J. Biol. Chem.,266:7214 (1991); Lee et al. J. Biol. Chem, 268: 12412 (1993); Yurewiczet al. Biochem. Biophys. Acta 1174:211 (1993)). Comparison of the humanZPA genomic structure with the human ZPB genomic structure shows theseregions to be confined to exons 12, 13, and 14 of the human ZPA gene andexons 5, 6, and 7 of the human ZPB gene. This suggests that thishomology might be due to a partial ancestral gene duplication. The ZPBproteins contain 21 conserved cysteine residues. The first 11 of thesedo not align with those in the ZPA proteins, but the last 10 match well.This extends the homology to approximately 270 amino acids, coveringexons 11-16 of the ZPA gene and exons 4-9 of the ZPB gene, although theoverall homology of the expanded region is slightly lower (approximately43%). The remainder of the ZPA and ZPB genes show very little homologywith each other, and the ZPC genes also show no extensive homology tothe ZPA genes. In addition, the ZPA gene has no extensive sequencesimilarity to non-ZP nucleic acid and protein sequences in Genbank andthe SwissProt data banks.

EXAMPLE 14 Comparison of ZPB DNA and of Deduced Amino Acid Sequences

Table 6 shows the comparison of the six known ZPB DNA and proteinsequences (the bovine and cynomolgus cDNA fragments are only compared tothe corresponding regions of the other full-length ZPB sequences).

                  TABLE 6                                                         ______________________________________                                        ZPB HOMOLOGY                                                                    PROTEIN HOMOLOGY                                                                                                   C.                                       Rabbit Bovine Porcine Feline Monkey Human                                   ______________________________________                                        Rabbit  --      75.3%    65.3% 60.1% 70.2% 65.2%                                Bovine 78.8% -- 82.3% 71.2% 69.9% 69.6%                                       Porcine 74.2% 86.2% -- 63.7% 63.6% 63.1%                                      Feline 69.5% 78.7% 72.9% -- 70.3% 64.6%                                       C. Monkey 78.9% 78.5% 78.2% 78.6% -- 92.3%                                    Human 74.3% 80.8% 73.3% 74.2%   95% --                                      ______________________________________                                                  DNA HOMOLOGY                                                    

The data are presented as cross-wise comparison of the ZPB protein andDNA sequences. The comparison of the protein sequences are shown in theupper right hand side of the table, above the diagonal dashed lines. Thecomparison of the DNA sequences are shown in the lower left hand side ofthe table, below the diagonal dashed lines.

The data shows considerable ZPB homology among members of differentmammalian species. As was the case with ZPA, this homology is mostpronounced between members of the same order within the class mammalia.For example, the human and cynomolgus monkey sequences (pnmata) and thepig and cow sequences (ungulata) have the most homology to each other.With only a few exceptions (noted below), the ZPB sequences show nohomology to other DNA or protein sequences in the GenBank or SwissProtdatabases. Hybridization experiments suggest that the ZPB transcriptsare ovary specific.

Comparisons of the deduced amino acid sequences of the ZPB clones showmore divergence within this genetic group than within the ZPA and ZPCgroups. Comparison of the rabbit ZPB and porcine ZPB shows the sequencesto be predominantly collinear (74% homologous) except that the rabbithas an additional upstream ATG codon which adds six codons to the rabbitsequence.

The feline ZPB sequence has two additional amino acid inserts, whichtotal 38 additional codons, in the first quarter of the gene, comparedto the porcine and rabbit sequences. Both inserts occur just aftercysteine residues, which suggests that if the cysteines are involved indisulfide bridges, these regions might form unique epitopes. However,the feline gene is still 73% homologous to porcine gene and 70%homologous to the rabbit gene.

The human gene has a sequence homologous to the first of the inserts inthe cat sequence, but not the second. However, there are consensussplice site donor and acceptor sequences adjacent to this extra regionin the human sequence, which if used would leave the coding sequence inframe. Therefore, the sequence representing exon 2 could actually be twosmall exons (122 and 103 bp) separated by a small intron (84 bp). Thiswould make the human sequence in this region identical to the pigsequence. The first extra region in the cat sequence is also flanked byin frame splice site donor and acceptor signals. If the extra region wasremoved from the cat sequence, it would differ from the pig sequence byonly a single amino acid. However, the cat sequence was obtained from acDNA clone made from an mRNA that appears to be fully processed. Thesecond extra region in the cat sequence does not contain in frame splicesite donor or acceptor signals, and therefore is probably not due to thepresence of an unprocessed intron.

The cynomolgus monkey and human sequences have an additional sevencodons at the C-terminus when compared to the other ZPB sequences. Inthe cynomolgus monkey, this is due to a two-base pair deletion, whichcauses a frameshift mutation which puts the termination codon used bythe other species out of frame. The human sequence also contains thisdeletion, but in addition, there is also a base change that eliminatesthis termination codon.

There are 21 conserved cysteine residues in the ZPB proteins, the final10 of which occur in a region that has homology to the ZPA proteins.This homology was noted previously (Schwoebel et al., supra; Lee et al.supra, 1993; Yurewicz et al. supra, 1993), but examination of thegenomic structure of the human ZPA and ZPB genes allowed the homology tobe extended to approximately 270 amino acids. This homology could be dueto a partial ancestral gene duplication. In addition to the conservedcysteine residues, the pig ZPB protein contains one additional cysteineresidue in the putative leader sequence, and the human sequence containsfour additional cysteine residues. The first of these is in the putativeleader sequence (in a different location than pig), the second is in theregion containing the additional insert, and the last two are in theC-terminal extension caused by the mutated termination codon. These lasttwo extra cysteine residues are conserved in the cynomolgus monkeysequence.

All of the ZP proteins contain a putative transmembrane domain near theC-terminus. However, the canonical furin proteolytic processing signal(R-X-R/K-R, Hosaka et al. supra, 1991), which occurs just prior to thetransmembrane domain in all of the ZPA and ZPC proteins, is altered inthe human (S-R-R-R), cynomolgus monkey (S-R-R-N) and rabbit (S-R-R-R)ZPB sequences. The significance of this is unknown, but it may indicatethat these proteins are processed by a related system with specificityfor di- or tribasic sequences, since the release of the putativetransmembrane domain would be necessary for the ZPB protein to move asthe ZP layer grows. There appears to be a great deal of proteolyticprocessing of the pig ZPA and ZPB (Yurewicz et al. supra,) proteins.There is no data concerning the post-translational modification of theZPB proteins of cat, cow, cynomolgus monkey or human. The physiologicsignificance of this processing is unknown, but differential processingwould present an avenue of variation among species of the highlyconserved ZP proteins.

There is a question of whether humans actually transcribe the ZPB gene.Since the amount of human ovarian mRNA recovered was so small, there wasnot enough RNA to both construct a cDNA library and perform a Northernanalysis. However, since cynomolgus monkey transcribes the ZPB gene, itis probable that the highly homologous human ZPB gene is alsotranscribed.

The apparent lack of a ZPB cDNA in the dog cDNA library is anotherpuzzle. All of the libraries screened which contained any zona pellucidagene contained all three genes, except the dog. However, mRNA isolatedfrom the ovary of a six-month old dog (the library was made from theovary of a four-month old dog), includes a ZPB mRNA that comigrates withthe porcine and cynomolgus monkey ZPB mRNA on a Northern blot. Onepossibility to explain the lack of a canine ZPB cDNA is that thetranscriptional timing of the three ZP genes is spread out, and sincethe ovary used to make the library was young, the transcription of theZPB gene occurs later than the ZPA and ZPC genes (Andersen and Simpson,1973).

EXAMPLE 15 Comparison of ZPC DNA and Deduced Amino Acid Sequences

Table 7 shows the comparison of the DNA and deduced amino acid sequencesfrom all of the ZPC cDNAs and genes.

                                      TABLE 7                                     __________________________________________________________________________    ZPC HOMOLOGY                                                                  PROTEIN HOMOLOGY                                                                   Mouse                                                                             Hamster                                                                           Rabbit                                                                            Pig Cow Dog Cat Monkey                                                                            Human                                    __________________________________________________________________________    Mouse                                                                              --  78.8%                                                                             65.9%                                                                             65.6%                                                                             64.0%                                                                             64.7%                                                                             63.3%                                                                             64.4%                                                                             67.0%                                      Hamster 84.7% -- 65.9% 65.6% 63.5% 65.1% 63.6% 68.2% 68.0%                    Rabbit 70.1% 71.3% -- 68.2% 68.5% 65.3% 64.1% 59.4% 68.5%                     Pig 71.5% 72.0% 74.6% -- 83.6% 75.7% 72.8% 69.2% 73.7%                        Cow 70.5% 71.4% 74.5% 86.5% -- 74.5% 72.8% 67.4% 71.1%                        Dog 70.1% 71.9% 71.5% 79.8% 80.3% -- 79.2% 66.5% 70.1%                        Cat 70.9% 71.6% 73.0% 79.3% 80.0% 84.3% -- 71.1% 70.5%                        Monkey 72.4% 74.1% 71.3% 76.6% 77.2% 73.8% 77.8% -- 90.6%                     Human 74.1% 75.0% 76.2% 80.0% 79.6% 77.7% 78.8% 94.4% --                    __________________________________________________________________________    DNA HOMOLOGY                                                              

The data are presented as a cross-wise comparison of the ZPC protein andDNA sequences. The comparison of the protein sequences are shown in theupper right hand side of the table, above the diagonal dashed lines. Thecomparison of the DNA sequences are shown in the lower left hand side ofthe table, below the diagonal dashed lines.

ZPC proteins and DNA sequences show a higher degree of homology than theZPA and ZPB DNAs and proteins. As was the case with ZPA and ZPB, thehomology is most pronounced in members of the same order within theclass mammalia; the human and cynomolgus monkey sequences (primata), thecat and dog sequences (carnivora), the pig and cow sequences (ungulata),and the mouse and hamster sequences (rodenta). The ZPC transcripts areovary specific, based on Northern blot analysis and comparison to thesequences in the GenBank and SwissProt databases detects no significantnon-ZP homology. Comparison of the deduced amino acid sequences of theknown ZPC genes detects three regions that contain large numbers ofnon-consensus sequences. These regions are: the putative leadersequences and the first 20-25 amino acids of the mature protein; theregion containing the peptide that was identified as a sperm-bindingregion in the mouse (Millar et al. Science 216:935-938 (1989)); and theC-terminal region of the proteins that might be removed from the matureprotein at the furin processing site (see below).

The epitope identified as a putative sperm-binding site (Millar et al.supra, 1989) occurs immediately before a furin proteolytic cleavage site(Hosaka et al., 1991). The furin site (R-X-R/K-R) is highly conserved inall of the ZPC sequences. However, it should be noted that the canineZPC sequence contains a second furin site, 19 amino acids upstream fromthe first furin site. Also as is the case with ZPA and ZPB, cleavage byfurin of the ZPC proteins would remove a putative membrane anchorsequence (Klein et al., 1985), which would allow the processed ZPCprotein to move toward the outer layer of the expanding oocyte.Therefore, this sperm-binding site probably represents the C-terminus ofthe mature proteins. However, there is very little homology (evenbetween hamster and mouse) in the regions of the ZPC proteinscorresponding to this epitope. This might indicate that this regioncontributes to the species specificity of sperm-egg binding.

The variation that is seen at the C-terminus of the ZPC proteins occursin the putative transmembrane region. This variation could indicate thatthis amino acid sequence is less important than the overallhydrophobicity of the amino acids in this region, similar to the lack ofhomology seen in leader sequences. However, it is also possible thatthis variation signifies a species-specific function for this region.

Each ZPC sequence contains 14 conserved cysteine residues, but eachsequence also has one or two extra cysteine residues that are sharedonly with one or a few other sequences. These extra cysteine residuesare near the N- or C-terminus of the proteins, where the greatestsequence variation exists. However, the large number of conservedcysteine residues probably indicates that the overall structure of thecentral core of all of these proteins is quite conserved.

EXAMPLE 16 Immunization of Cynomolgus Monkeys With HSPZ

A sexually mature cynomolgus monkey was immunized with HSPZ to test theability of HSPZ to induce infertility. HSPZ was prepared as described inExample 6. HSPZ was mixed with the following GMDP/oil adjuvant. 50 μgGMDP (N-acetyl-D-glucosaminyl-(β1-4)-N-acetylmuramyl-D-isoglutamine)(CC. Biotech, Poway, Calif.); 42.1 of mineral oil, 15.8% pluronic VC-121(block polymer polyols, BASF-Wyandotte, Parsippany, N.J.). The animalreceived a series of 4 subcutaneous injections of 1 mg of HSPZ in theGMDP/oil adjuvant beginning with a priming dose followed four weekslater by a booster dose, which was followed by two booster doses fiveweeks apart which were followed six weeks later by a final dose. Thisdosage regimen resulted in an anovulatory monkey having antibody titersagainst its cynomolgus monkey heat-solubilized zona pellucida preparedas described for HSPZ. The peak antibody titers to cynomolgus monkeyHSPZ were 1:8000-1:16,000.

A fractionated preparation of HSPZ which is essentially native porcineZPA and ZPB was prepared by isoelectric focusing, as described inExample 6 and was used to vaccinate cynomolgus monkeys using 1 mg offractionated HSPZ in GMDP/oil injected subcutaneously according to thefollowing schedule: a priming dose was given followed approximately 6weeks later by a booster dose followed by a final booster dose 11 weeksafter the previous booster dose. The immunized monkeys achieved peakantibody titers of 1:4,000-1:8,000 against monkey heat-solubilized zonapellucida while maintaining a regular ovulatory cycle. However, despitemaintaining a regular ovulatory cycle, the monkeys remained infertileuntil their antibody titers to monkey heat-solubilized zona pellucidafell below 1:500 after which the animals became pregnant upon breeding.

Immunization of cynomolgus monkeys with recombinant baculovirus producedcanine ZPC and porcine ZPC (prepared as described in Example 18) failedto induce infertility despite inducing antibody production againstmonkey heat-solubilized zona pellucida. One possible explanation forthis is that the glycosylation pattern of ZP proteins produced in thebaculovirus system may prevent recognition of the epitopes responsiblefor induction of infertility.

Bacterially produced porcine ZPA, ZPB, and ZPC described aboveadministered to cynomolgus monkeys failed to induce detectable antibodytiters against cynomolgus monkey heat-solubilized zona pellucida eventhough antibody titers against the presented antigens were produced.

EXAMPLE 17 Mapping of Mammalian Zona Pellucida Protein Epitopes

A Pin Technology™ Epitope Scanning Kit purchased from Chiron MimotopesU.S., Emeryville, Calif. (Catalog No. PT-02-20000A) was used for mappingepitopes in Zona Pellucida proteins. The procedures described in the kitmanual were followed, with the exception of modifications in the ELISAtesting procedure (described below).

Briefly, Pin Technology software was installed in a United BusinessMachines 486/33 computer according to the manufacturer's instructions.The protein sequence was entered into the computer program, the desiredpeptide length, and degree of overlap between peptides were selected,and a protocol containing the daily requirements of activated protectedamino acid derivatives and their location in the coupling tray wells wasprinted. Prior to use, the pins were first washed once withdimethylformamide (DMF), and then with methanol three times, each washlasting for two minutes. The pin block was air dried and the pins weredeprotected by agitation in a 20% mixture of piperidine in DMF at roomtemperature for 30 minutes. The pins were washed again as describedabove, except that the washes were for 5 minutes each, and the pin blockwas then air dried. The required amino acid derivative solutions wereprepared and dispensed into the wells of the synthesis tray according tothe protocol for the current cycle. The dried mimotope pins were washedonce more in a DMF bath for 5 minutes and then positioned appropriatelyin the wells of the synthesis tray. The assembly was then sealed in aplastic bag and incubated at 30° C. for approximately 22 hours. On thefollowing day, the pin block was removed from the coupling tray andsubjected to the same cycle of washing, deprotection, and coupling stepsas outlined above; however, using the amino acid derivatives and theirtray location appropriate to the next cycle. The foregoing cycle ofwashing, deprotection, washing, and coupling was repeated until thepeptide sequences were completed.

After coupling the terminal amino acids of the peptides, the pin blockwas washed, air dried, deprotected, washed and air dried as before. Theterminal amino groups of the peptides were then acetylated by immersionof the pins in a mixture containing 5 parts DMF, 2 parts aceticanhydride, and 1 part triethylamine, by volume, dispensed in the wellsof a polypropylene coupling tray, and incubating at 30° C. for 90minutes. The pin block was removed, subjected to another washingsequence as before, and air dried.

Side chain deprotection of the peptides was performed by agitating thepin block in a mixture containing 95 parts trifluoroacetic acid, 2.5parts anisole, and 2.5 parts ethanedithiol, by volume, at roomtemperature for 4 hours. The pin block was then air dried forapproximately 10 minutes, sonicated in a bath containing 0. 1%hydrochloric acid in a mixture containing equal parts of methanol anddeionized water, by volume, for 15 minutes, and finally air dried.

Prior to ELISA testing, the pins were subjected to a disruptionprocedure involving sonication in a bath consisting of a mixturecontaining 39 parts sodium dihydrogen orthophosphate, 25 parts sodiumdodecyl sulfate, 0.1 part 2-mercaptoethanol, and 2500 parts deionizedwater, by weight, adjusted to pH 7.2 with 50% sodium hydroxide solution.The sonication was performed at 55 to 60° C. for approximately 45minutes. The pin block was then washed by immersion with gentleagitation in three sequential baths of deionized water at 60 degrees forthree minutes each. Finally, the pin block was immersed in gentlyboiling methanol for approximately 4 minutes and then air dried.

Preparation of Antisera

Antisera directed against zona pellucida proteins was prepared byimmunizing the appropriate animals with the appropriate zona pellucidaprotein using procedures well known in the art and described in E.Harlow and D. Lane in Antibodies, A Laboratory Manual, Chapter 5, ColdSpring Harbor Laboratory, 1988 which is incorporated herein byreference. Biotinylated antisera was prepared by a modification of theprocedure described in Harlow supra (page 314). Briefly, to a solutioncontaining between 1 and 3 mg per ml of the selected antibody IgGfraction in phosphate buffer with saline (PBS) at pH 7.2 was added asolution containing 25 to 250 micrograms biotinamidocaproate,N-hydroxysuccinimide ester (Sigma, Cat No. B2643) in dimethyl sulfoxideat a concentration of 10 mg/ml. The mixture was mixed well and thenincubated at room temperature for 4 hours. One molar ammonium chloridesolution in the amount corresponding to 20 microliters per 250micrograms biotin ester was added, and the resulting mixture wasincubated at room temperature for 10 minutes. Unreacted biotin ester wasthen removed by extensive diafiltration with PBS using a Centricon-30(TM) microconcentrator devices (Amicon Division, W. R. Grace & Co.,Inc., Beverly Mass.). The dilution factor for the resulting conjugatewas determined by ELISA titration against the appropriate nativeprotein.

ELISA Testing

A modification of the procedure described in the Epitope Scanning Kitmanual was employed.

After disruption, the mimotope pins were blocked by incubation with"supercocktail" (10 g albumin, 10 g bovine serum albumin, and 1 ml Tween20 detergent per liter of PBS) at room temperature for 1 hour. This wasfollowed by incubation at room temperature for 2 hours withappropriately diluted biotinylated antisera. The pins were washed 4times with PBS containing 0.5% Tween 20 (PBST) at room temperature for10 minutes each time, with agitation.

The pins were then incubated at room temperature for 1 hour with thesecondary antibody, horseradish peroxidase-streptavidin conjugate (ZymedLaboratories, Inc., South San Francisco, Calif.) diluted 1:2500 withPBST. They were washed again as described above.

Substrate buffer was prepared by combining 200 ml 1.0 M. disodiumhydrogen orthophosphate solution with 160 ml 1.0 M. citric acidsolution, diluting the mixture with 1640 ml deionized water, andadjusting to pH 4.0 using either citric acid or sodium hydroxidesolutions. Substrate solution was prepared by dissolving 10 mg2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt in20 ml substrate buffer and adding 6 microliters 30% hydrogen peroxide.The mimotope pins were incubated at room temperature with this solution,using microtiter plates containing 150 microliters per well. When colordevelopment appeared to be appropriate for measurement by an ELISA platereader, the pin block was removed and the plate was read at a wavelengthof 450 nm. The pin block was then disrupted by the procedure describedabove.

The data were entered into the Pin Technology™ computer program, whichperformed statistical analysis and evaluation and furnished a print-outof the results identifying the strongest binding epitopes. Briefly, the25% of the wells having the lowest optical density readings were assumedto represent background in each experiment. The mean value and thestandard deviation of these readings were calculated. Significantrecognition of peptides by antisera was attributed to the pinscorresponding to those wells showing absorbance readings greater thanthe sum of the background mean and three standard deviations from themean.

Human ZPA epitopes were examined for reactivity with mouse anti-human ZPantiserum prepared as described above. Peptides of 15 amino acids inlength were synthesized beginning with amino acid number 1 asillustrated in SEQ ID NO. 43. Successive peptides having a 7-amino acidoverlap with the preceding peptide of the series were synthesized. Thefollowing peptides were shown to bind mouse anti-human ZP antiserum:1-15, 9-23, 25-39, 33-47, 65-79, 81-95, 89-103, 97-111, 105-119,113-127, 121-135, 129-143, 145-159, 153-167, 161-175, 193-207, 209-223,217-231, 225-239, 241-255, 249-263, 273-287, 281-295, 289-303, 305-319,313-327, 321-335, 329-343, 337-351, 345-359, 385-399, 393-407, 401-415,409-423, 417-431, 425-439, 441-455, 449-463, 457-471, 481-495, 489-503,497-511, 505-519, 513-527, 521-535, 537-551, 545-559, 561-575, 569-583,577-591, 585-599, 601-615, 609-623, 617-631, 625-639, 633-647, 641-655,665-679, 697-711, 705-719, 713-727, 721-735, and 729-743.

Similarly, human ZPB epitopes were mapped using mouse anti-human ZPantiserum. In these experiments, 15 amino acid peptides were synthesizedbeginning with amino acid number 1 as set out in SEQ ID NO. 41. Theoverlap between successive peptides in this case was 9 amino acids. Thefollowing peptides were shown to bind mouse anti-human ZP antiserum:7-21, 25-39, 31-45, 49-63, 67-81, 73-87, 79-93, 91-105, 103-117,121-135, 193-207, 205-219, 211-225, 217-231, 223-237, 229-243, 253-267,259-273, 265-279, 283-297, 289-303, 295-309, 301-315, 307-321, 313-327,319-333, 343-357, 349-363, 355-369, 367-381, 373-387, 379-393, 385-399,403-417, 409-423, 415-429, 421-435, 433-447, 439-453, 445-459, 451-465,481-495, 487-501, 499-513, 505-519, 511-525, 523-537, 529-543, and547-561.

Human ZPC epitopes were mapped using mouse anti-human ZP antiserum. Inthese experiments, the 15 amino acid peptides were synthesized beginningwith amino acid number 1 as set out in Chamberlin et al., Proc. Nat'lAcad. Sci. USA 87:6014-6018 (1990) which is incorporated herein byreference. The overlap between successive peptides was 10 amino acids.The following peptides were shown to bind mouse anti-human ZP antiserum:21-35, 51-65, 116-130, 146-160, 151-165, 181-195, 241-255, 251-265,271-285, 296-310, 321-335, 401-415, and 411-425.

Canine ZPC epitopes were mapped using rabbit anti-canine ZP antiserum.In these experiments, the 15 amino acid peptides were synthesizedbeginning at amino acid number 1 set out in SEQ ID NO. 10. The overlapbetween successive peptides was 5 amino acids. The following peptideswere shown to bind rabbit anti-canine ZP antiserum: 51-65, 61-75, 81-95,131-145, 181-195, and 301-315.

Feline ZPC epitopes were mapped using rabbit anti-feline ZP antiserum.In these experiments, the 15 amino acid peptides were synthesizedbeginning at amino acid number 1 as set out in SEQ ID NO. 18. Theoverlap between successive peptides was 5 amino acids. The followingpeptides were shown to bind rabbit anti-feline ZP: 36-50, 46-60, 56-70,76-90, 96-110, 106-120, 116-130, 126-140, 136-150, 146-160, 156-170,186-200, 196-210, 246-260, 266-280, 276-290, 286-300, 296-310, 316-330,326-340, 336-350, 346-360, 376-390, 396-410, and 406-420.

Bovine ZPC epitopes were mapped using rabbit anti-bovine ZP antiserum.In these experiments, the overlapping 15 amino acid peptides weresynthesized beginning at amino acid number 1 as set out in SEQ ID NO.24. The overlap between peptides was 10 amino acids. The followingpeptides were shown to be reactive with rabbit anti-bovine ZP antiserum:1-15, 31-45, 51-65, 56-70, 61-75, 76-90, 106-120, 111-125, 116-130,121-135, 131-145, 136-150, 141-155, 146-160, 151-165, 161-175, 181-195,186-200, 191-205, 196-210, 201-215, 206-220, 216-230, 226-240, 241-255,246-260, 261-275, 266-280, 271-285, 276-290, 291-305, 296-310, 301-315,316-330, 321-335, 326-340, 331-345, 336-350, 341-355, 356-370, 361-375,376-390, 381-395, 386-400, 396-410, 401-415, and 406-420.

EXAMPLE 18 Immunization of Dogs with Recombinant ZPC Proteins

Dogs were immunized with various preparations of recombinant canine ZPC.The plasmid pZ169 bacterial expression vector (FIG. 5) was constructedas follows. The parent vector pZ98 (described in Example 9) was digestedwith the restriction enzymes Pvul and Bam HI, and the large fragment wasgel purified. Into this vector was ligated a fragment created byannealing the following oligonucleotides:

5' CGCCCTCCCAGCAACTGCACCATCACCACCATGGG 3' (SEQ ID NO.50); and

5' GATCCCCATGGTGGTGGTGATGGTGCAGTTGCTGGGAAGGGCGAT 3' (SEQ ID NO.51).

These oligonucleotides create a fragment with PvuI and BamHI ends, andcodes for the hexapeptide sequence His₆. This intermediate vector wasdigested with the restriction enzymes BamHI and EcoRI, and the largefragment was gel purified. Into this vector was ligated a fragmentcreated by annealing the following oligonucleotides:

5' GATCCCTCGAGCCACCATCACCACCATCATG 3' (SEQ ID NO.52); and

5' AATRCATGATGGTGGTGATGGTGGCTCGAGG 3' (SEQ ID NO.53).

These oligonucleotides create a fragment with BamHI and EcoRI ends andan XhoI site just downstream of the BamHI site, and which codes for thehexapeptide sequence His₆. This new vector was named pZ88, and containsunique BamHI and XhoI cloning sites between two His₆ sequences. Tocreate pZ169, the pZ88 vector was digested with the restriction enzymesBamHI and XhoI, and the large fragment was gel purified. Into thisvector was ligated a fragment generated by performing a PCR (polymerasechain reaction) of the canine ZPC cDNA using the followingoligonucleotides:

5' CCCGGATCCGCAGACCATCTGGCCAACTGAG 3' (SEQ ID NO.54); and

5' GCGCTCGAGGGCATATGGCTGCCAGTGTG 3' (SEQ ID NO.55).

This PCR creates a fragment containing amino acids 23-207 of the canineZPC sequence, with BamHI and XhoI ends. This new vector is named pZ169,(FIG. 5) and produces a protein containing amino acids 1-56 of the E.coli β-galactosidase sequence, His₆, amino acids 23-207 of the canineZPC sequence, His₆, and amino acids 1006-1023 of the E. coliβ-galactosidase sequence. This protein is referred to as N-terminalcanine ZPC. In FIG. 5, pTAC refers to the tac promoter described above;AmpR refers to an ampicillin resistance marker, ori is an E. coli originof replication sequences and pLacI is the lacI promoter which drivesexpression of the laci gene.

Recombinant canine ZPC was produced and purified as described in Example9. A baculovirus expression vector pZ145 was constructed as follows. Theparent vector pBlueBac2 (purchased from Invitrogen Corporation, SanDiego, Calif.) was digested with the restriction enzymes NheI and BamHI,and the large fragment was gel purified. Into this vector was ligated afragment generated by a PCR of the porcine ZPC cDNA using the followingoligonucleotide:

5' CGCGCTAGCAGATCTATGGCGCCGAGCTGGAGGTTC 3' (SEQ ID NO. 56); and

5' CGCGGATCCTATTAATGGTGGTGATGGTGGTGACTAGTGGACCCTTCCA 3' (SEQ ID NO.57).

This PCR creates a fragment with NheI and BamHI ends, and contains aminoacids 27-350 of the porcine ZPC sequence followed by an SpeI site andthe hexapeptide His₆. This new vector is named pZ147. To create thepZ145 vector, pZ147 is digested with NheI and SpeI and the largefragment is gel purified (this removes the pig ZPC sequence). Into thisvector was ligated a fragment generated by a PCR of the canine ZPC cDNAusing the following oligonucleotides:

5' CCCGCTAGCAGATCTATGGGGCTGAGCTATGGAATTTTC 3' (SEQ ID NO.58); and

5' CGCACTAGTTGACCCCTCTATACCATGATCACTA 3' (SEQ ID NO.59).

This PCR creates a fragment with NheI and SpeI ends, and contains aminoacids 1-379 of the canine sequence. Transformants of this ligation werescreened for the presence of the inserted NheI/SpeI fragment in thecorrect orientation (since the NheI and SpeI sticky ends are identical).This new vector is named pZ145, (FIG. 6) and produces a proteincontaining amino acids 1-379 of the DZPC sequence followed by His₆. Thisprotein is referred to as baculo-canine ZPC. In FIG. 6, pP representsthe baculovirus polyhedrin promoter, AmpR represents an ampicillinresistance marker, LacZ represents the gene for β-galactosidase, pE is aconstituitive promoter which drives the expression of LacZ and ori isthe E. coli origin of replication.

Recombinant baculovirus derived canine ZPC was produced byco-transfecting insect SF9 cells with pZ145 and Autographica californicamultiply enveloped nuclear polyhedrosis virus (AcMNPV) using methodswell known in the art as described in the MAXBAC™ kit purchased fromInvitrogen, San Diego, Calif. Recombinant canine ZPC produced in SF9cells was prepared from cotransfected SF9 cells as follows.Cotransfected cells were harvested and pelleted by centrifugation andrecombinant canine ZPC was purified as was described in Example 9 forpurification from a cell pellet. Recombinant canine ZPC may also beisolated from the culture medium and purified on a Ni-column asdescribed in Example 9.

Other expression vectors which are capable of expressing zona pellucidaencoding nucleotide sequences under the control of a variety ofregulatory sequences are within the scope of the present invention andare readily constructed using methods well known in the art.

Recombinant zona pellucida proteins may also be modified to increasetheir potential antigenicity by a variety of methods well known in theart. For example, a recombinant dog ZPC was modified by palmitylationwas prepared as follows. Approximately 1 mg of recombinant ZPC producedusing the plasmid pZ169 as described above was brought to a finalconcentration of 8M urea (total volume 0.2-0.3 mis.). A palmitylationsolution (Pl₂ O/TEA) was then prepared by adding palmitic anhydride totriethylamine to give a final concentration of palmitic anhydride of 20mg/ml of triethylamine.

Approximately 10 μl of Pl₂ O/TEA solution was added to 1 mg ofrecombinant canine ZPC in 8M urea (described above). The mixture wasallowed to stand at room temperature for a least two hours after whichthe preparation was ready for mixture with GMDP/oil adjuvant.

Chitosan modification is another useful modification of canine ZPC forthe practice of the present invention. Briefly, 1.5 ml of sterilemineral oil was added to 1.5 ml of recombinant canine ZPC solutionprepared as described above using the plasmid pZ169 (2 mg/ml ZPC, 3 mgtotal is 8M urea) was mixed with 5 drops of Arlacel A (mannidemonooleate, Sigma, St, Louis, Mo.). Subsequently, 0.75 ml of Chitosan(2% wt/vol. is 0.5M sodium acetate, pH 5.0) was added, and the mixturewas sonicated for 10-20 seconds, followed by the addition of 0.045 ml of50% NaOH and another round of sonication for 10-20 seconds. Finally, 10μl of 10 mg/ml GMDP/8M urea was added.

A group of three dogs was immunized five times each at one-monthintervals with subcutaneous injections of 1 mg doses of the N-terminalcanine ZPC modified by the addition of chitosan prepared as describedabove. Immunized dogs developed antibody titers of 1:8000-1:16000against heat solubilized dog zona pellucida (self-titers) using methodsdescribed above. The estrus cycle of the dogs showing self-titers wasanovulatory and prolonged (4-6 weeks instead of the normal 10-day to14-day cycle for normal dogs). Of the three immunized dogs, two haveexperienced their first estrus; one of the two dogs exhibited estrus sixmonths after the first immunization and was bred and found to beinfertile. The second of the two dogs experienced estrus and remainedinfertile nine months after the first immunization. The third dog hasyet to experience estrus more than nine months after immunization.

Another group of four dogs were immunized three times at one-monthintervals using 1 mg doses of palmitylated canine ZPC (prepared asdescribed above) in GMDP/oil adjuvant administered subcutaneously. Theseanimals achieved self-titers (against heat solubilized dog zonapellucida) of 1:4000-1:8000. Nearly seven months after immunization, twoof the four dogs experienced estrus and remain infertile. The remainingtwo dogs have yet to experience estrus.

Another set of dogs was immunized 3 times at one-month intervals, usingsubcutaneous injections of 1 mg of recombinant canine ZPC produced usingpZ166, (a plasmid similar to pZ169 but containing a DNA sequenceencoding amino acids 23-379 of the canine ZPC protein) in GMDP/oiladjuvant. These animals failed to develop self-titers and becamepregnant after breeding. Similarly, dogs immunized with canine ZPCfragments produced using the baculovirus system failed to induceinfertility.

EXAMPLE 19 Vaccination of Cows and Cats with Recombinant Zona PellucidaProteins

Preliminary studies were undertaken to assess the ability of recombinantzona pellucida proteins to induce infertility in cows and cats.

Cows were injected with 3 or more doses (in GMDP (250 μg) oil adjuvant)of 1 mg of a variety of recombinantly derived ZPC proteins from canineand porcine sources including canine ZPC produced using the plasmidpZ169 as shown in FIG. 5. Recombinant proteins were administered in anunmodified form and in palmitylated and chitosan modified forms. None ofthe ZP protein preparations induced self-titers or infertility in thevaccinated cows. Further studies are underway using differentrecombinant preparations of zona pellucida proteins and differing dosageregimens in attempts to induce self-titers and infertility in cows.

Similarly, cats were vaccinated with the following recombinant zonapellucida proteins: a mixture of recombinant feline ZPA, ZPB, and ZPC;porcine ZPC produced using pZ156 as described above and shown in FIG. 3;and canine ZPC produced using the plasmid pZ169 described above andshown in FIG. 5. Cats vaccinated using these ZP protein preparationsproduced antibody to the vaccine proteins, but produced no self-titersand were consequently fertile. Studies are ongoing to determine theeffects of modifying the recombinant zona pellucida proteins in attemptsto stimulate the production of self-titers and to induce infertility.

Studies are also ongoing to select other recombinantly derived zonapellucida protein fragments for testing as possibleimmunocontraceptives.

Numerous modifications in variations in the practice of the invention asillustrated in the above examples are expected to occur to those ofordinary skill in the art. Consequently, the illustrative examples arenot intended to limit the scope of the invention as set out in theappended claims.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                   - -  - - (1) GENERAL INFORMATION:                                             - -    (iii) NUMBER OF SEQUENCES: 59                                          - -  - - (2) INFORMATION FOR SEQ ID NO:1:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2214 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Sus scrof - #a                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: sig.sub.-- - #peptide                                           (B) LOCATION: 12..119                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 120..2153                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 12..2153                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                               - - GAATTCCGGG C AGG CAC AGA GGA GAC AGT GGG AGA - #CCC TTA AGC TGG        CTC      50                                                                                  Arg His Arg - #Gly Asp Ser Gly Arg Pro Leu Ser Trp Leu                       -36 -35   - #              -30   - #              -25             - - AGT GCA AGC TGG AGG TCA CTT CTT CTA TTT TT - #C CCC CTT GTG ACT TCA           98                                                                       Ser Ala Ser Trp Arg Ser Leu Leu Leu Phe Ph - #e Pro Leu Val Thr Ser                       -20      - #           -15      - #           -10                  - - GTG AAC TCC ATA GGT GTC AAT CAG TTG GTG AA - #T ACT GCC TTC CCA GGT          146                                                                       Val Asn Ser Ile Gly Val Asn Gln Leu Val As - #n Thr Ala Phe Pro Gly                    -5         - #          1        - #       5                          - - ATT GTC ACT TGC CAT GAA AAT AGA ATG GTA GT - #G GAA TTT CCA AGA ATT          194                                                                       Ile Val Thr Cys His Glu Asn Arg Met Val Va - #l Glu Phe Pro Arg Ile            10                 - # 15                 - # 20                 - # 25       - - CTT GGC ACT AAG ATA CAG TAC ACC TCT GTG GT - #G GAC CCT CTT GGT CTT          242                                                                       Leu Gly Thr Lys Ile Gln Tyr Thr Ser Val Va - #l Asp Pro Leu Gly Leu                            30 - #                 35 - #                 40              - - GAA ATG ATG AAC TGT ACT TAT GTT CTG GAC CC - #A GAA AAC CTC ACC CTG          290                                                                       Glu Met Met Asn Cys Thr Tyr Val Leu Asp Pr - #o Glu Asn Leu Thr Leu                        45     - #             50     - #             55                  - - AAG GCC CCA TAT GAA GCC TGT ACC AAA AGA GT - #G CGT GGC CAT CAC CAA          338                                                                       Lys Ala Pro Tyr Glu Ala Cys Thr Lys Arg Va - #l Arg Gly His His Gln                    60         - #         65         - #         70                      - - ATG ACC ATC AGA CTC ATA GAT GAC AAT GCT GC - #T TTA AGA CAA GAG GCT          386                                                                       Met Thr Ile Arg Leu Ile Asp Asp Asn Ala Al - #a Leu Arg Gln Glu Ala                75             - #     80             - #     85                          - - CTC ATG TAT CAC ATC AGC TGT CCT GTT ATG GG - #A GCA GAA GGC CCT GAT          434                                                                       Leu Met Tyr His Ile Ser Cys Pro Val Met Gl - #y Ala Glu Gly Pro Asp            90                 - # 95                 - #100                 - #105       - - CAG CAT TCG GGA TCC ACA ATC TGC ATG AAA GA - #T TTC ATG TCT TTT ACC          482                                                                       Gln His Ser Gly Ser Thr Ile Cys Met Lys As - #p Phe Met Ser Phe Thr                           110  - #               115  - #               120              - - TTT AAC TTT TTT CCC GGG ATG GCT GAC GAA AA - #T GTG AAA CGT GAG GAT          530                                                                       Phe Asn Phe Phe Pro Gly Met Ala Asp Glu As - #n Val Lys Arg Glu Asp                       125      - #           130      - #           135                  - - TCG AAG CAG CGC ATG GGA TGG AGC CTT GTA GT - #T GGT GAC GGT GAA AGA          578                                                                       Ser Lys Gln Arg Met Gly Trp Ser Leu Val Va - #l Gly Asp Gly Glu Arg                   140          - #       145          - #       150                      - - GCC CGA ACT CTG ACC TTT CAG GAG GCC ATG AC - #C CAA GGA TAT AAT TTC          626                                                                       Ala Arg Thr Leu Thr Phe Gln Glu Ala Met Th - #r Gln Gly Tyr Asn Phe               155              - #   160              - #   165                          - - CTG ATA GAG AAC CAG AAG ATG AAC ATC CAA GT - #G TCA TTC CAT GCC ACT          674                                                                       Leu Ile Glu Asn Gln Lys Met Asn Ile Gln Va - #l Ser Phe His Ala Thr           170                 1 - #75                 1 - #80                 1 -      #85                                                                              - - GGA GTG ACT CGC TAC TCG CAA GGT AAC AGT CA - #T CTC TAC ATG GTA        CCT      722                                                                    Gly Val Thr Arg Tyr Ser Gln Gly Asn Ser Hi - #s Leu Tyr Met Val Pro                          190  - #               195  - #               200              - - CTG AAG CTT AAA CAT GTA TCT CAT GGG CAG TC - #T CTC ATC TTA GCA TCA          770                                                                       Leu Lys Leu Lys His Val Ser His Gly Gln Se - #r Leu Ile Leu Ala Ser                       205      - #           210      - #           215                  - - CAA CTC ATC TGT GTG GCA GAT CCT GTG ACC TG - #T AAT GCC ACA CAC GTG          818                                                                       Gln Leu Ile Cys Val Ala Asp Pro Val Thr Cy - #s Asn Ala Thr His Val                   220          - #       225          - #       230                      - - ACT CTT GCC ATA CCA GAG TTT CCT GGG AAG CT - #A AAA TCC GTG AAC TTG          866                                                                       Thr Leu Ala Ile Pro Glu Phe Pro Gly Lys Le - #u Lys Ser Val Asn Leu               235              - #   240              - #   245                          - - GGA AGT GGG AAT ATT GCT GTG AGC CAG CTG CA - #C AAA CAC GGG ATT GAA          914                                                                       Gly Ser Gly Asn Ile Ala Val Ser Gln Leu Hi - #s Lys His Gly Ile Glu           250                 2 - #55                 2 - #60                 2 -      #65                                                                              - - ATG GAA ACA ACA AAC GGC CTG AGG TTG CAT TT - #C AAC CAA ACT CTT        CTC      962                                                                    Met Glu Thr Thr Asn Gly Leu Arg Leu His Ph - #e Asn Gln Thr Leu Leu                          270  - #               275  - #               280              - - AAA ACA AAT GTC TCT GAA AAA TGC CTA CCA CA - #T CAG TTG TAC TTA TCT         1010                                                                       Lys Thr Asn Val Ser Glu Lys Cys Leu Pro Hi - #s Gln Leu Tyr Leu Ser                       285      - #           290      - #           295                  - - TCA CTC AAG CTG ACT TTT CAC AGT CAA CTA GA - #G GCA GTA TCC ATG GTG         1058                                                                       Ser Leu Lys Leu Thr Phe His Ser Gln Leu Gl - #u Ala Val Ser Met Val                   300          - #       305          - #       310                      - - ATT TAT CCT GAG TGT CTC TGT GAG TCA ACA GT - #C TCT TTA GTT TCA GAG         1106                                                                       Ile Tyr Pro Glu Cys Leu Cys Glu Ser Thr Va - #l Ser Leu Val Ser Glu               315              - #   320              - #   325                          - - GAG CTA TGC ACT CAG GAT GGG TTT ATG GAC GT - #C AAG GTC CAC AGC CAC         1154                                                                       Glu Leu Cys Thr Gln Asp Gly Phe Met Asp Va - #l Lys Val His Ser His           330                 3 - #35                 3 - #40                 3 -      #45                                                                              - - CAA ACA AAA CCA GCT CTC AAC TTG GAT ACC CT - #C AGG GTG GGA GAC        TCA     1202                                                                    Gln Thr Lys Pro Ala Leu Asn Leu Asp Thr Le - #u Arg Val Gly Asp Ser                          350  - #               355  - #               360              - - TCC TGC CAG CCA ACC TTT AAA GCT CCA GCT CA - #G GGG CTG GTA CAG TTT         1250                                                                       Ser Cys Gln Pro Thr Phe Lys Ala Pro Ala Gl - #n Gly Leu Val Gln Phe                       365      - #           370      - #           375                  - - CGC ATA CCC CTG AAT GGA TGT GGA ACA AGA CA - #T AAG TTC AAG AAT GAC         1298                                                                       Arg Ile Pro Leu Asn Gly Cys Gly Thr Arg Hi - #s Lys Phe Lys Asn Asp                   380          - #       385          - #       390                      - - AAA GTC ATC TAT GAA AAT GAA ATA CAT GCT CT - #C TGG GCA GAT CCT CCA         1346                                                                       Lys Val Ile Tyr Glu Asn Glu Ile His Ala Le - #u Trp Ala Asp Pro Pro               395              - #   400              - #   405                          - - AGC GCC GTT TCC AGA GAT AGT GAG TTC AGA AT - #G ACA GTG AGG TGC TCT         1394                                                                       Ser Ala Val Ser Arg Asp Ser Glu Phe Arg Me - #t Thr Val Arg Cys Ser           410                 4 - #15                 4 - #20                 4 -      #25                                                                              - - TAC AGC AGC AGC AAC ATG CTA ATA AAT ACC AA - #T GTT GAA AGT CTT        CCT     1442                                                                    Tyr Ser Ser Ser Asn Met Leu Ile Asn Thr As - #n Val Glu Ser Leu Pro                          430  - #               435  - #               440              - - TCT CCA GAG GCC TCA GTG AAG CCA GGT CCA CT - #T ACC CTG ACT CTG CAA         1490                                                                       Ser Pro Glu Ala Ser Val Lys Pro Gly Pro Le - #u Thr Leu Thr Leu Gln                       445      - #           450      - #           455                  - - ACC TAC CCA GAT AAC GCC TAC CTG CAG CCT TA - #T GGG GAC AAG GAG TAC         1538                                                                       Thr Tyr Pro Asp Asn Ala Tyr Leu Gln Pro Ty - #r Gly Asp Lys Glu Tyr                   460          - #       465          - #       470                      - - CCT GTG GTG AAA TAT CTC CGC CAA CCA ATT TA - #C CTA GAA GTG AGA ATC         1586                                                                       Pro Val Val Lys Tyr Leu Arg Gln Pro Ile Ty - #r Leu Glu Val Arg Ile               475              - #   480              - #   485                          - - CTC AAC AGG ACT GAC CCC AAC ATC AAG CTG GT - #C TTG GAT GAC TGC TGG         1634                                                                       Leu Asn Arg Thr Asp Pro Asn Ile Lys Leu Va - #l Leu Asp Asp Cys Trp           490                 4 - #95                 5 - #00                 5 -      #05                                                                              - - GCA ACA TCC ACA GAG GAC CCA GCC TCT CTC CC - #C CAG TGG AAT GTT        GTC     1682                                                                    Ala Thr Ser Thr Glu Asp Pro Ala Ser Leu Pr - #o Gln Trp Asn Val Val                          510  - #               515  - #               520              - - ATG GAT GGC TGT GAA TAC AAC CTG GAC AAC CA - #C AGA ACC ACC TTC CAT         1730                                                                       Met Asp Gly Cys Glu Tyr Asn Leu Asp Asn Hi - #s Arg Thr Thr Phe His                       525      - #           530      - #           535                  - - CCG GTG GGC TCC TCC GTG ACC TAT CCT AAC CA - #C CAT CAG AGG TTT GAT         1778                                                                       Pro Val Gly Ser Ser Val Thr Tyr Pro Asn Hi - #s His Gln Arg Phe Asp                   540          - #       545          - #       550                      - - GTG AAG ACC TTT GCC TTT GTG TCA GGG GCC CA - #A GGG GTC TCT CAA CTG         1826                                                                       Val Lys Thr Phe Ala Phe Val Ser Gly Ala Gl - #n Gly Val Ser Gln Leu               555              - #   560              - #   565                          - - GTC TAC TTC CAC TGC AGT GTC TTC ATC TGC AA - #T CAA CTC TCT CCC ACC         1874                                                                       Val Tyr Phe His Cys Ser Val Phe Ile Cys As - #n Gln Leu Ser Pro Thr           570                 5 - #75                 5 - #80                 5 -      #85                                                                              - - TTC TCT CTG TGT TCT GTG ACT TGC CAT GGG CC - #A TCT AGG AGC CGG        CGA     1922                                                                    Phe Ser Leu Cys Ser Val Thr Cys His Gly Pr - #o Ser Arg Ser Arg Arg                          590  - #               595  - #               600              - - GCT ACA GGG ACC ACT GAG GAA GAG AAA ATG AT - #A GTG AGT CTC CCG GGC         1970                                                                       Ala Thr Gly Thr Thr Glu Glu Glu Lys Met Il - #e Val Ser Leu Pro Gly                       605      - #           610      - #           615                  - - CCC ATC CTG CTG TTG TCA GAT GGC TCT TCA CT - #C AGA GAT GCT GTG AAC         2018                                                                       Pro Ile Leu Leu Leu Ser Asp Gly Ser Ser Le - #u Arg Asp Ala Val Asn                   620          - #       625          - #       630                      - - TCT AAA GGA TCC AGA ACC AAC GGA TAT GTT GC - #T TTT AAA ACT ATG GTT         2066                                                                       Ser Lys Gly Ser Arg Thr Asn Gly Tyr Val Al - #a Phe Lys Thr Met Val               635              - #   640              - #   645                          - - GCT ATG GTT GCT TCA GCA GGC ATC GTG GCA AC - #T CTA GGC CTC ATC AGC         2114                                                                       Ala Met Val Ala Ser Ala Gly Ile Val Ala Th - #r Leu Gly Leu Ile Ser           650                 6 - #55                 6 - #60                 6 -      #65                                                                              - - TAC CTG CAC AAA AAA AGA ATC ATG ATG TTA AA - #T CAC TAATTTGGAT              2160                                                                      Tyr Leu His Lys Lys Arg Ile Met Met Leu As - #n His                                           670  - #               675                                     - - TTTCAAATAA AAGTGGAAGT AAGCCTCTTC TAAAAAAAAA AAAAACCGGA AT - #TC             2214                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:2:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 713 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                               - - Arg His Arg Gly Asp Ser Gly Arg Pro Leu Se - #r Trp Leu Ser Ala Ser     36 -35                - # -30                - # -25                            - - Trp Arg Ser Leu Leu Leu Phe Phe Pro Leu Va - #l Thr Ser Val Asn Ser     20                 - - #15                 - - #10                  - #-5       - - Ile Gly Val Asn Gln Leu Val Asn Thr Ala Ph - #e Pro Gly Ile Val Thr                        - #1               5   - #               10                  - - Cys His Glu Asn Arg Met Val Val Glu Phe Pr - #o Arg Ile Leu Gly Thr               15         - #         20         - #         25                      - - Lys Ile Gln Tyr Thr Ser Val Val Asp Pro Le - #u Gly Leu Glu Met Met           30             - #     35             - #     40                          - - Asn Cys Thr Tyr Val Leu Asp Pro Glu Asn Le - #u Thr Leu Lys Ala Pro       45                 - # 50                 - # 55                 - # 60       - - Tyr Glu Ala Cys Thr Lys Arg Val Arg Gly Hi - #s His Gln Met Thr Ile                       65 - #                 70 - #                 75              - - Arg Leu Ile Asp Asp Asn Ala Ala Leu Arg Gl - #n Glu Ala Leu Met Tyr                   80     - #             85     - #             90                  - - His Ile Ser Cys Pro Val Met Gly Ala Glu Gl - #y Pro Asp Gln His Ser               95         - #        100         - #        105                      - - Gly Ser Thr Ile Cys Met Lys Asp Phe Met Se - #r Phe Thr Phe Asn Phe          110              - #   115              - #   120                          - - Phe Pro Gly Met Ala Asp Glu Asn Val Lys Ar - #g Glu Asp Ser Lys Gln      125                 1 - #30                 1 - #35                 1 -      #40                                                                              - - Arg Met Gly Trp Ser Leu Val Val Gly Asp Gl - #y Glu Arg Ala Arg        Thr                                                                                             145  - #               150  - #               155             - - Leu Thr Phe Gln Glu Ala Met Thr Gln Gly Ty - #r Asn Phe Leu Ile Glu                  160      - #           165      - #           170                  - - Asn Gln Lys Met Asn Ile Gln Val Ser Phe Hi - #s Ala Thr Gly Val Thr              175          - #       180          - #       185                      - - Arg Tyr Ser Gln Gly Asn Ser His Leu Tyr Me - #t Val Pro Leu Lys Leu          190              - #   195              - #   200                          - - Lys His Val Ser His Gly Gln Ser Leu Ile Le - #u Ala Ser Gln Leu Ile      205                 2 - #10                 2 - #15                 2 -      #20                                                                              - - Cys Val Ala Asp Pro Val Thr Cys Asn Ala Th - #r His Val Thr Leu        Ala                                                                                             225  - #               230  - #               235             - - Ile Pro Glu Phe Pro Gly Lys Leu Lys Ser Va - #l Asn Leu Gly Ser Gly                  240      - #           245      - #           250                  - - Asn Ile Ala Val Ser Gln Leu His Lys His Gl - #y Ile Glu Met Glu Thr              255          - #       260          - #       265                      - - Thr Asn Gly Leu Arg Leu His Phe Asn Gln Th - #r Leu Leu Lys Thr Asn          270              - #   275              - #   280                          - - Val Ser Glu Lys Cys Leu Pro His Gln Leu Ty - #r Leu Ser Ser Leu Lys      285                 2 - #90                 2 - #95                 3 -      #00                                                                              - - Leu Thr Phe His Ser Gln Leu Glu Ala Val Se - #r Met Val Ile Tyr        Pro                                                                                             305  - #               310  - #               315             - - Glu Cys Leu Cys Glu Ser Thr Val Ser Leu Va - #l Ser Glu Glu Leu Cys                  320      - #           325      - #           330                  - - Thr Gln Asp Gly Phe Met Asp Val Lys Val Hi - #s Ser His Gln Thr Lys              335          - #       340          - #       345                      - - Pro Ala Leu Asn Leu Asp Thr Leu Arg Val Gl - #y Asp Ser Ser Cys Gln          350              - #   355              - #   360                          - - Pro Thr Phe Lys Ala Pro Ala Gln Gly Leu Va - #l Gln Phe Arg Ile Pro      365                 3 - #70                 3 - #75                 3 -      #80                                                                              - - Leu Asn Gly Cys Gly Thr Arg His Lys Phe Ly - #s Asn Asp Lys Val        Ile                                                                                             385  - #               390  - #               395             - - Tyr Glu Asn Glu Ile His Ala Leu Trp Ala As - #p Pro Pro Ser Ala Val                  400      - #           405      - #           410                  - - Ser Arg Asp Ser Glu Phe Arg Met Thr Val Ar - #g Cys Ser Tyr Ser Ser              415          - #       420          - #       425                      - - Ser Asn Met Leu Ile Asn Thr Asn Val Glu Se - #r Leu Pro Ser Pro Glu          430              - #   435              - #   440                          - - Ala Ser Val Lys Pro Gly Pro Leu Thr Leu Th - #r Leu Gln Thr Tyr Pro      445                 4 - #50                 4 - #55                 4 -      #60                                                                              - - Asp Asn Ala Tyr Leu Gln Pro Tyr Gly Asp Ly - #s Glu Tyr Pro Val        Val                                                                                             465  - #               470  - #               475             - - Lys Tyr Leu Arg Gln Pro Ile Tyr Leu Glu Va - #l Arg Ile Leu Asn Arg                  480      - #           485      - #           490                  - - Thr Asp Pro Asn Ile Lys Leu Val Leu Asp As - #p Cys Trp Ala Thr Ser              495          - #       500          - #       505                      - - Thr Glu Asp Pro Ala Ser Leu Pro Gln Trp As - #n Val Val Met Asp Gly          510              - #   515              - #   520                          - - Cys Glu Tyr Asn Leu Asp Asn His Arg Thr Th - #r Phe His Pro Val Gly      525                 5 - #30                 5 - #35                 5 -      #40                                                                              - - Ser Ser Val Thr Tyr Pro Asn His His Gln Ar - #g Phe Asp Val Lys        Thr                                                                                             545  - #               550  - #               555             - - Phe Ala Phe Val Ser Gly Ala Gln Gly Val Se - #r Gln Leu Val Tyr Phe                  560      - #           565      - #           570                  - - His Cys Ser Val Phe Ile Cys Asn Gln Leu Se - #r Pro Thr Phe Ser Leu              575          - #       580          - #       585                      - - Cys Ser Val Thr Cys His Gly Pro Ser Arg Se - #r Arg Arg Ala Thr Gly          590              - #   595              - #   600                          - - Thr Thr Glu Glu Glu Lys Met Ile Val Ser Le - #u Pro Gly Pro Ile Leu      605                 6 - #10                 6 - #15                 6 -      #20                                                                              - - Leu Leu Ser Asp Gly Ser Ser Leu Arg Asp Al - #a Val Asn Ser Lys        Gly                                                                                             625  - #               630  - #               635             - - Ser Arg Thr Asn Gly Tyr Val Ala Phe Lys Th - #r Met Val Ala Met Val                  640      - #           645      - #           650                  - - Ala Ser Ala Gly Ile Val Ala Thr Leu Gly Le - #u Ile Ser Tyr Leu His              655          - #       660          - #       665                      - - Lys Lys Arg Ile Met Met Leu Asn His                                          670              - #   675                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:3:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1699 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Sus scrof - #a                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: sig.sub.-- - #peptide                                           (B) LOCATION: 38..445                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 446..1648                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 38..1648                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                               - - GAATTCCGGG TGGAAGTACC TGTTCTCCGC AGGCGCT ATG TGG TTG - #CGG CCG TCC          55                                                                                          - #                  - #     Met Trp Leu Arg Pro Ser                          - #                  - #     -136-135                        - - ATC TGG CTC TGC TTT CCG CTG TGT CTT GCT CT - #G CCA GGC CAG TCT CAG          103                                                                       Ile Trp Leu Cys Phe Pro Leu Cys Leu Ala Le - #u Pro Gly Gln Ser Gln          130                -125 - #                -120 - #               -115          - - CCC AAA GCA GCA GAT GAC CTT GGT GGC CTC TA - #C TGT GGG CCA AGC AGC          151                                                                       Pro Lys Ala Ala Asp Asp Leu Gly Gly Leu Ty - #r Cys Gly Pro Ser Ser                           -110 - #               -105  - #              -100             - - TTT CAT TTC TCC ATA AAT CTT CTC AGC CAG GA - #C ACA GCA ACT CCT CCT          199                                                                       Phe His Phe Ser Ile Asn Leu Leu Ser Gln As - #p Thr Ala Thr Pro Pro                       -95      - #           -90      - #           -85                  - - GCA CTG GTG GTT TGG GAC AGG CGC GGG CGG CT - #G CAC AAG CTG CAG AAT          247                                                                       Ala Leu Val Val Trp Asp Arg Arg Gly Arg Le - #u His Lys Leu Gln Asn                   -80          - #       -75          - #       -70                      - - GAC TCT GGC TGT GGC ACG TGG GTC CAC AAG GG - #C CCA GGC AGC TCC ATG          295                                                                       Asp Ser Gly Cys Gly Thr Trp Val His Lys Gl - #y Pro Gly Ser Ser Met               -65              - #   -60              - #   -55                          - - GGA GTG GAA GCA TCC TAC AGA GGC TGC TAT GT - #G ACT GAG TGG GAC TCT          343                                                                       Gly Val Glu Ala Ser Tyr Arg Gly Cys Tyr Va - #l Thr Glu Trp Asp Ser          50                 - - #45                 - - #40                 - -        #35                                                                              - - CAC TAC CTC ATG CCC ATT GGA CTT GAA GAA GC - #A GAT GCA GGT GGA        CAC      391                                                                    His Tyr Leu Met Pro Ile Gly Leu Glu Glu Al - #a Asp Ala Gly Gly His                          -30  - #               -25  - #               -20              - - AGA ACA GTC ACA GAG ACG AAA CTG TTT AAG TG - #C CCT GTG GAT TTC CTA          439                                                                       Arg Thr Val Thr Glu Thr Lys Leu Phe Lys Cy - #s Pro Val Asp Phe Leu                       -15      - #           -10      - #            -5                  - - GCT CTT GAT GTT CCA ACC ATT GGC CTT TGT GA - #T GCT GTC CCA GTG TGG          487                                                                       Ala Leu Asp Val Pro Thr Ile Gly Leu Cys As - #p Ala Val Pro Val Trp                     1        - #       5           - #       10                          - - GAC CGA TTG CCA TGT GCT CCT CCA CCC ATC AC - #T CAA GGA GAA TGC AAG          535                                                                       Asp Arg Leu Pro Cys Ala Pro Pro Pro Ile Th - #r Gln Gly Glu Cys Lys            15                 - # 20                 - # 25                 - # 30       - - CAG CTT GGC TGC TGC TAC AAC TCG GAA GAG GT - #C CCT TCT TGT TAC TAT          583                                                                       Gln Leu Gly Cys Cys Tyr Asn Ser Glu Glu Va - #l Pro Ser Cys Tyr Tyr                            35 - #                 40 - #                 45              - - GGA AAC ACA GTG ACC TCA CGC TGT ACC CAA GA - #T GGC CAC TTC TCC ATC          631                                                                       Gly Asn Thr Val Thr Ser Arg Cys Thr Gln As - #p Gly His Phe Ser Ile                        50     - #             55     - #             60                  - - GCT GTG TCT CGC AAT GTG ACC TCA CCT CCA CT - #G CTC TGG GAT TCT GTG          679                                                                       Ala Val Ser Arg Asn Val Thr Ser Pro Pro Le - #u Leu Trp Asp Ser Val                    65         - #         70         - #         75                      - - CAC CTG GCC TTC AGA AAT GAC AGT GAA TGT AA - #A CCT GTG ATG GAA ACA          727                                                                       His Leu Ala Phe Arg Asn Asp Ser Glu Cys Ly - #s Pro Val Met Glu Thr                80             - #     85             - #     90                          - - CAC ACT TTT GTC CTC TTC CGG TTT CCA TTT AG - #T TCC TGT GGG ACT GCA          775                                                                       His Thr Phe Val Leu Phe Arg Phe Pro Phe Se - #r Ser Cys Gly Thr Ala            95                 - #100                 - #105                 - #110       - - AAA CGG GTA ACT GGG AAC CAG GCG GTA TAT GA - #A AAT GAG CTG GTA GCA          823                                                                       Lys Arg Val Thr Gly Asn Gln Ala Val Tyr Gl - #u Asn Glu Leu Val Ala                           115  - #               120  - #               125              - - GCT CGG GAT GTG AGG ACT TGG AGC CAT GGT TC - #T ATT ACC CGA GAC AGC          871                                                                       Ala Arg Asp Val Arg Thr Trp Ser His Gly Se - #r Ile Thr Arg Asp Ser                       130      - #           135      - #           140                  - - ATC TTC AGG CTT CGA GTC AGT TGT ATC TAC TC - #T GTA AGT AGC AGT GCT          919                                                                       Ile Phe Arg Leu Arg Val Ser Cys Ile Tyr Se - #r Val Ser Ser Ser Ala                   145          - #       150          - #       155                      - - CTC CCA GTT AAC ATC CAG GTT TTC ACT CTC CC - #A CCA CCG CTT CCG GAG          967                                                                       Leu Pro Val Asn Ile Gln Val Phe Thr Leu Pr - #o Pro Pro Leu Pro Glu               160              - #   165              - #   170                          - - ACC CAC CCT GGA CCT CTT ACT CTG GAG CTT CA - #G ATT GCC AAA GAT GAA         1015                                                                       Thr His Pro Gly Pro Leu Thr Leu Glu Leu Gl - #n Ile Ala Lys Asp Glu           175                 1 - #80                 1 - #85                 1 -      #90                                                                              - - CGC TAT GGC TCC TAC TAC AAT GCT AGT GAC TA - #C CCG GTG GTG AAA        TTG     1063                                                                    Arg Tyr Gly Ser Tyr Tyr Asn Ala Ser Asp Ty - #r Pro Val Val Lys Leu                          195  - #               200  - #               205              - - CTT CGG GAG CCC ATC TAT GTG GAG GTC TCT AT - #C CGT CAC CGA ACA GAC         1111                                                                       Leu Arg Glu Pro Ile Tyr Val Glu Val Ser Il - #e Arg His Arg Thr Asp                       210      - #           215      - #           220                  - - CCC AGT CTC GGG CTG CAC CTG CAC CAG TGC TG - #G GCC ACA CCC GGC ATG         1159                                                                       Pro Ser Leu Gly Leu His Leu His Gln Cys Tr - #p Ala Thr Pro Gly Met                   225          - #       230          - #       235                      - - AGC CCC CTG CTC CAG CCA CAG TGG CCC ATG CT - #A GTC AAT GGA TGC CCC         1207                                                                       Ser Pro Leu Leu Gln Pro Gln Trp Pro Met Le - #u Val Asn Gly Cys Pro               240              - #   245              - #   250                          - - TAC ACT GGA GAC AAC TAC CAG ACC AAA CTG AT - #C CCT GTC CAG AAA GCC         1255                                                                       Tyr Thr Gly Asp Asn Tyr Gln Thr Lys Leu Il - #e Pro Val Gln Lys Ala           255                 2 - #60                 2 - #65                 2 -      #70                                                                              - - TCA AAC CTG CTA TTT CCT TCT CAC TAC CAG CG - #T TTC AGT GTT TCC        ACC     1303                                                                    Ser Asn Leu Leu Phe Pro Ser His Tyr Gln Ar - #g Phe Ser Val Ser Thr                          275  - #               280  - #               285              - - TTC AGT TTT GTG GAC TCT GTG GCA AAG CAG GC - #A CTC AAG GGA CCG GTG         1351                                                                       Phe Ser Phe Val Asp Ser Val Ala Lys Gln Al - #a Leu Lys Gly Pro Val                       290      - #           295      - #           300                  - - TAT CTG CAT TGT ACT GCA TCG GTC TGC AAG CC - #T GCA GGG GCA CCG ATC         1399                                                                       Tyr Leu His Cys Thr Ala Ser Val Cys Lys Pr - #o Ala Gly Ala Pro Ile                   305          - #       310          - #       315                      - - TGT GTG ACA ACC TGT CCT GCT GCC AGA CGA AG - #A AGA AGT TCT GAC ATC         1447                                                                       Cys Val Thr Thr Cys Pro Ala Ala Arg Arg Ar - #g Arg Ser Ser Asp Ile               320              - #   325              - #   330                          - - CAT TTT CAG AAT GGC ACT GCT AGC ATT TCT AG - #C AAG GGT CCC ATG ATT         1495                                                                       His Phe Gln Asn Gly Thr Ala Ser Ile Ser Se - #r Lys Gly Pro Met Ile           335                 3 - #40                 3 - #45                 3 -      #50                                                                              - - CTA CTC CAA GCC ACT CGG GAC TCT TCA GAA AG - #G CTC CAT AAA TAC        TCA     1543                                                                    Leu Leu Gln Ala Thr Arg Asp Ser Ser Glu Ar - #g Leu His Lys Tyr Ser                          355  - #               360  - #               365              - - AGG CCT CCT GTA GAC TCC CAT GCT CTG TGG GT - #G GCT GGC CTC TTG GGA         1591                                                                       Arg Pro Pro Val Asp Ser His Ala Leu Trp Va - #l Ala Gly Leu Leu Gly                       370      - #           375      - #           380                  - - AGC TTA ATT ATT GGA GCC TTG TTA GTG TCC TA - #C CTG GTC TTC AGG AAA         1639                                                                       Ser Leu Ile Ile Gly Ala Leu Leu Val Ser Ty - #r Leu Val Phe Arg Lys                   385          - #       390          - #       395                      - - TGG AGA TGAGTTACTC AGACCAAATG TGTCAATAAA ACCAATAAAA CA - #AAACCGGA          1695                                                                       Trp Arg                                                                           400                                                                        - - ATTC                 - #                  - #                  - #               1699                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:4:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 536 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                               - - Met Trp Leu Arg Pro Ser Ile Trp Leu Cys Ph - #e Pro Leu Cys Leu Ala     136 -135               - #-130                -12 - #5                          - - Leu Pro Gly Gln Ser Gln Pro Lys Ala Ala As - #p Asp Leu Gly Gly Leu     120                -115 - #                -110 - #               -105          - - Tyr Cys Gly Pro Ser Ser Phe His Phe Ser Il - #e Asn Leu Leu Ser Gln                      -100 - #                -95 - #                -90             - - Asp Thr Ala Thr Pro Pro Ala Leu Val Val Tr - #p Asp Arg Arg Gly Arg                  -85      - #           -80      - #           -75                  - - Leu His Lys Leu Gln Asn Asp Ser Gly Cys Gl - #y Thr Trp Val His Lys              -70          - #       -65          - #       -60                      - - Gly Pro Gly Ser Ser Met Gly Val Glu Ala Se - #r Tyr Arg Gly Cys Tyr          -55              - #   -50              - #   -45                          - - Val Thr Glu Trp Asp Ser His Tyr Leu Met Pr - #o Ile Gly Leu Glu Glu     40                 - - #35                 - - #30                 - -        #25                                                                              - - Ala Asp Ala Gly Gly His Arg Thr Val Thr Gl - #u Thr Lys Leu Phe        Lys                                                                                             -20  - #               -15  - #               -10             - - Cys Pro Val Asp Phe Leu Ala Leu Asp Val Pr - #o Thr Ile Gly Leu Cys                   -5     - #              1    - #           5                      - - Asp Ala Val Pro Val Trp Asp Arg Leu Pro Cy - #s Ala Pro Pro Pro Ile           10             - #     15             - #     20                          - - Thr Gln Gly Glu Cys Lys Gln Leu Gly Cys Cy - #s Tyr Asn Ser Glu Glu       25                 - # 30                 - # 35                 - # 40       - - Val Pro Ser Cys Tyr Tyr Gly Asn Thr Val Th - #r Ser Arg Cys Thr Gln                       45 - #                 50 - #                 55              - - Asp Gly His Phe Ser Ile Ala Val Ser Arg As - #n Val Thr Ser Pro Pro                   60     - #             65     - #             70                  - - Leu Leu Trp Asp Ser Val His Leu Ala Phe Ar - #g Asn Asp Ser Glu Cys               75         - #         80         - #         85                      - - Lys Pro Val Met Glu Thr His Thr Phe Val Le - #u Phe Arg Phe Pro Phe           90             - #     95             - #    100                          - - Ser Ser Cys Gly Thr Ala Lys Arg Val Thr Gl - #y Asn Gln Ala Val Tyr      105                 1 - #10                 1 - #15                 1 -      #20                                                                              - - Glu Asn Glu Leu Val Ala Ala Arg Asp Val Ar - #g Thr Trp Ser His        Gly                                                                                             125  - #               130  - #               135             - - Ser Ile Thr Arg Asp Ser Ile Phe Arg Leu Ar - #g Val Ser Cys Ile Tyr                  140      - #           145      - #           150                  - - Ser Val Ser Ser Ser Ala Leu Pro Val Asn Il - #e Gln Val Phe Thr Leu              155          - #       160          - #       165                      - - Pro Pro Pro Leu Pro Glu Thr His Pro Gly Pr - #o Leu Thr Leu Glu Leu          170              - #   175              - #   180                          - - Gln Ile Ala Lys Asp Glu Arg Tyr Gly Ser Ty - #r Tyr Asn Ala Ser Asp      185                 1 - #90                 1 - #95                 2 -      #00                                                                              - - Tyr Pro Val Val Lys Leu Leu Arg Glu Pro Il - #e Tyr Val Glu Val        Ser                                                                                             205  - #               210  - #               215             - - Ile Arg His Arg Thr Asp Pro Ser Leu Gly Le - #u His Leu His Gln Cys                  220      - #           225      - #           230                  - - Trp Ala Thr Pro Gly Met Ser Pro Leu Leu Gl - #n Pro Gln Trp Pro Met              235          - #       240          - #       245                      - - Leu Val Asn Gly Cys Pro Tyr Thr Gly Asp As - #n Tyr Gln Thr Lys Leu          250              - #   255              - #   260                          - - Ile Pro Val Gln Lys Ala Ser Asn Leu Leu Ph - #e Pro Ser His Tyr Gln      265                 2 - #70                 2 - #75                 2 -      #80                                                                              - - Arg Phe Ser Val Ser Thr Phe Ser Phe Val As - #p Ser Val Ala Lys        Gln                                                                                             285  - #               290  - #               295             - - Ala Leu Lys Gly Pro Val Tyr Leu His Cys Th - #r Ala Ser Val Cys Lys                  300      - #           305      - #           310                  - - Pro Ala Gly Ala Pro Ile Cys Val Thr Thr Cy - #s Pro Ala Ala Arg Arg              315          - #       320          - #       325                      - - Arg Arg Ser Ser Asp Ile His Phe Gln Asn Gl - #y Thr Ala Ser Ile Ser          330              - #   335              - #   340                          - - Ser Lys Gly Pro Met Ile Leu Leu Gln Ala Th - #r Arg Asp Ser Ser Glu      345                 3 - #50                 3 - #55                 3 -      #60                                                                              - - Arg Leu His Lys Tyr Ser Arg Pro Pro Val As - #p Ser His Ala Leu        Trp                                                                                             365  - #               370  - #               375             - - Val Ala Gly Leu Leu Gly Ser Leu Ile Ile Gl - #y Ala Leu Leu Val Ser                  380      - #           385      - #           390                  - - Tyr Leu Val Phe Arg Lys Trp Arg                                                  395          - #       400                                             - -  - - (2) INFORMATION FOR SEQ ID NO:5:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1326 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Sus scrof - #a                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: sig.sub.-- - #peptide                                           (B) LOCATION: 25..105                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 106..1290                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 25..1290                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                               - - GAATTCCGGG GCCTTGTGAG TGCC ATG GCG CCG AGC TGG A - #GG TTC TTC GTC            51                                                                                         - #         Met Ala Pro Ser Trp - #Arg Phe Phe Val                            - #         -27     -25   - #              -20               - - TGC TTT CTG CTC TGG GGA GGT ACA GAG CTA TG - #C AGC CCG CAG CCC GTC           99                                                                       Cys Phe Leu Leu Trp Gly Gly Thr Glu Leu Cy - #s Ser Pro Gln Pro Val                       -15      - #           -10      - #            -5                  - - TGG CAG GAC GAA GGC CAG CGC TTG AGG CCC TC - #A AAG CCA CCC ACC GTA          147                                                                       Trp Gln Asp Glu Gly Gln Arg Leu Arg Pro Se - #r Lys Pro Pro Thr Val                     1        - #       5           - #       10                          - - ATG GTG GAG TGT CAG GAG GCC CAG CTG GTG GT - #C ATT GTC AGC AAA GAC          195                                                                       Met Val Glu Cys Gln Glu Ala Gln Leu Val Va - #l Ile Val Ser Lys Asp            15                 - # 20                 - # 25                 - # 30       - - CTT TTC GGT ACC GGG AAG CTC ATC AGG CCT GC - #A GAT CTC AGC CTG GGC          243                                                                       Leu Phe Gly Thr Gly Lys Leu Ile Arg Pro Al - #a Asp Leu Ser Leu Gly                            35 - #                 40 - #                 45              - - CCT GCA AAG TGT GAG CCG CTG GTC TCT CAG GA - #C ACG GAC GCA GTG GTC          291                                                                       Pro Ala Lys Cys Glu Pro Leu Val Ser Gln As - #p Thr Asp Ala Val Val           50                  - #55                  - #60                               - - AGG TTT GAG GTT GGG CTG CAC GAG TGT GGC AG - #C AGC TTG CAG GTG ACT          339                                                                       Arg Phe Glu Val Gly Leu His Glu Cys Gly Se - #r Ser Leu Gln Val Thr                    65         - #         70         - #         75                      - - GAT GAT GCT CTG GTG TAC AGC ACC TTC CTG CG - #C CAT GAC CCC CGC CCT          387                                                                       Asp Asp Ala Leu Val Tyr Ser Thr Phe Leu Ar - #g His Asp Pro Arg Pro                80             - #     85             - #     90                          - - GCA GGA AAC CTG TCC ATC CTG AGG ACG AAC CG - #T GCG GAG GTC CCC ATC          435                                                                       Ala Gly Asn Leu Ser Ile Leu Arg Thr Asn Ar - #g Ala Glu Val Pro Ile            95                 - #100                 - #105                 - #110       - - GAG TGT CAC TAC CCC AGG CAG GGC AAC GTG AG - #C AGC TGG GCC ATC CTG          483                                                                       Glu Cys His Tyr Pro Arg Gln Gly Asn Val Se - #r Ser Trp Ala Ile Leu                           115  - #               120  - #               125              - - CCC ACC TGG GTG CCC TTC AGG ACC ACG GTG TT - #C TCC GAG GAG AAG CTG          531                                                                       Pro Thr Trp Val Pro Phe Arg Thr Thr Val Ph - #e Ser Glu Glu Lys Leu                       130      - #           135      - #           140                  - - GTG TTC TCT CTG CGC CTG ATG GAG GAA AAC TG - #G AGT GCC GAG AAG ATG          579                                                                       Val Phe Ser Leu Arg Leu Met Glu Glu Asn Tr - #p Ser Ala Glu Lys Met                   145          - #       150          - #       155                      - - ACG CCC ACC TTC CAG CTG GGG GAC AGA GCC CA - #C CTC CAG GCC CAA GTC          627                                                                       Thr Pro Thr Phe Gln Leu Gly Asp Arg Ala Hi - #s Leu Gln Ala Gln Val               160              - #   165              - #   170                          - - CAC ACC GGC AGC CAC GTG CCA CTG AGG CTG TT - #T GTG GAC CAC TGT GTG          675                                                                       His Thr Gly Ser His Val Pro Leu Arg Leu Ph - #e Val Asp His Cys Val           175                 1 - #80                 1 - #85                 1 -      #90                                                                              - - GCC ACG CTG ACG CCG GAC TGG AAC ACC TCC CC - #C TCT CAC ACC ATC        GTG      723                                                                    Ala Thr Leu Thr Pro Asp Trp Asn Thr Ser Pr - #o Ser His Thr Ile Val                          195  - #               200  - #               205              - - GAC TTC CAC GGC TGT CTC GTG GAC GGT CTC AC - #T GAG GCC TCA TCT GCT          771                                                                       Asp Phe His Gly Cys Leu Val Asp Gly Leu Th - #r Glu Ala Ser Ser Ala                       210      - #           215      - #           220                  - - TTC AAA GCA CCT AGA CCT GGA CCA GAG ACG CT - #C CAG TTC ACC GTG GAT          819                                                                       Phe Lys Ala Pro Arg Pro Gly Pro Glu Thr Le - #u Gln Phe Thr Val Asp                   225          - #       230          - #       235                      - - GTG TTC CAT TTT GCT AAT GAT TCC AGA AAC AC - #G ATC TAC ATC ACC TGC          867                                                                       Val Phe His Phe Ala Asn Asp Ser Arg Asn Th - #r Ile Tyr Ile Thr Cys               240              - #   245              - #   250                          - - CAT CTG AAG GTC ACT CCG GCT GAC CGA GTC CC - #G GAC CAA CTC AAC AAA          915                                                                       His Leu Lys Val Thr Pro Ala Asp Arg Val Pr - #o Asp Gln Leu Asn Lys           255                 2 - #60                 2 - #65                 2 -      #70                                                                              - - GCC TGT TCC TTC AGC AAG TCC TCC AAC AGG TG - #G TCC CCG GTG GAA        GGG      963                                                                    Ala Cys Ser Phe Ser Lys Ser Ser Asn Arg Tr - #p Ser Pro Val Glu Gly                          275  - #               280  - #               285              - - CCT GCT GTT ATC TGT CGT TGC TGT CAC AAG GG - #G CAG TGT GGT ACC CCA         1011                                                                       Pro Ala Val Ile Cys Arg Cys Cys His Lys Gl - #y Gln Cys Gly Thr Pro                       290      - #           295      - #           300                  - - AGC CTT TCC AGG AAG CTG TCT ATG CCG AAG AG - #A CAG TCT GCT CCC CGC         1059                                                                       Ser Leu Ser Arg Lys Leu Ser Met Pro Lys Ar - #g Gln Ser Ala Pro Arg                   305          - #       310          - #       315                      - - AGT CGC AGG CAC GTG ACA GAT GAA GCA GAT GT - #C ACA GTG GGG CCT CTG         1107                                                                       Ser Arg Arg His Val Thr Asp Glu Ala Asp Va - #l Thr Val Gly Pro Leu               320              - #   325              - #   330                          - - ATC TTC CTG GGC AAG ACG AGT GAC CAC GGT GT - #G GAA GGG TCC ACC TCC         1155                                                                       Ile Phe Leu Gly Lys Thr Ser Asp His Gly Va - #l Glu Gly Ser Thr Ser           335                 3 - #40                 3 - #45                 3 -      #50                                                                              - - TCC CCC ACC TCG GTG ATG GTG GGC TTG GGC CT - #G GCC ACC GTG GTG        ACC     1203                                                                    Ser Pro Thr Ser Val Met Val Gly Leu Gly Le - #u Ala Thr Val Val Thr                          355  - #               360  - #               365              - - TTG ACT CTG GCT ACC ATT GTC CTG GGT GTG CC - #C AGG AGG CGT CGG GCT         1251                                                                       Leu Thr Leu Ala Thr Ile Val Leu Gly Val Pr - #o Arg Arg Arg Arg Ala                       370      - #           375      - #           380                  - - GCT GCC CAC CTT GTG TGC CCC GTG TCT GCT TC - #C CAA TAAAAGGAGA              1297                                                                       Ala Ala His Leu Val Cys Pro Val Ser Ala Se - #r Gln                                   385          - #       390                                             - - AACATGAAAA AAAAAAAAAA CCGGAATTC         - #                  - #              1326                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:6:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 421 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                               - - Met Ala Pro Ser Trp Arg Phe Phe Val Cys Ph - #e Leu Leu Trp Gly Gly     27     -25            - #     -20            - #     -15                        - - Thr Glu Leu Cys Ser Pro Gln Pro Val Trp Gl - #n Asp Glu Gly Gln Arg          -10              - #    -5              - #     1             - #  5       - - Leu Arg Pro Ser Lys Pro Pro Thr Val Met Va - #l Glu Cys Gln Glu Ala                       10 - #                 15 - #                 20              - - Gln Leu Val Val Ile Val Ser Lys Asp Leu Ph - #e Gly Thr Gly Lys Leu                   25     - #             30     - #             35                  - - Ile Arg Pro Ala Asp Leu Ser Leu Gly Pro Al - #a Lys Cys Glu Pro Leu               40         - #         45         - #         50                      - - Val Ser Gln Asp Thr Asp Ala Val Val Arg Ph - #e Glu Val Gly Leu His           55             - #     60             - #     65                          - - Glu Cys Gly Ser Ser Leu Gln Val Thr Asp As - #p Ala Leu Val Tyr Ser       70                 - # 75                 - # 80                 - # 85       - - Thr Phe Leu Arg His Asp Pro Arg Pro Ala Gl - #y Asn Leu Ser Ile Leu                       90 - #                 95 - #                100              - - Arg Thr Asn Arg Ala Glu Val Pro Ile Glu Cy - #s His Tyr Pro Arg Gln                  105      - #           110      - #           115                  - - Gly Asn Val Ser Ser Trp Ala Ile Leu Pro Th - #r Trp Val Pro Phe Arg              120          - #       125          - #       130                      - - Thr Thr Val Phe Ser Glu Glu Lys Leu Val Ph - #e Ser Leu Arg Leu Met          135              - #   140              - #   145                          - - Glu Glu Asn Trp Ser Ala Glu Lys Met Thr Pr - #o Thr Phe Gln Leu Gly      150                 1 - #55                 1 - #60                 1 -      #65                                                                              - - Asp Arg Ala His Leu Gln Ala Gln Val His Th - #r Gly Ser His Val        Pro                                                                                             170  - #               175  - #               180             - - Leu Arg Leu Phe Val Asp His Cys Val Ala Th - #r Leu Thr Pro Asp Trp                  185      - #           190      - #           195                  - - Asn Thr Ser Pro Ser His Thr Ile Val Asp Ph - #e His Gly Cys Leu Val              200          - #       205          - #       210                      - - Asp Gly Leu Thr Glu Ala Ser Ser Ala Phe Ly - #s Ala Pro Arg Pro Gly          215              - #   220              - #   225                          - - Pro Glu Thr Leu Gln Phe Thr Val Asp Val Ph - #e His Phe Ala Asn Asp      230                 2 - #35                 2 - #40                 2 -      #45                                                                              - - Ser Arg Asn Thr Ile Tyr Ile Thr Cys His Le - #u Lys Val Thr Pro        Ala                                                                                             250  - #               255  - #               260             - - Asp Arg Val Pro Asp Gln Leu Asn Lys Ala Cy - #s Ser Phe Ser Lys Ser                  265      - #           270      - #           275                  - - Ser Asn Arg Trp Ser Pro Val Glu Gly Pro Al - #a Val Ile Cys Arg Cys              280          - #       285          - #       290                      - - Cys His Lys Gly Gln Cys Gly Thr Pro Ser Le - #u Ser Arg Lys Leu Ser          295              - #   300              - #   305                          - - Met Pro Lys Arg Gln Ser Ala Pro Arg Ser Ar - #g Arg His Val Thr Asp      310                 3 - #15                 3 - #20                 3 -      #25                                                                              - - Glu Ala Asp Val Thr Val Gly Pro Leu Ile Ph - #e Leu Gly Lys Thr        Ser                                                                                             330  - #               335  - #               340             - - Asp His Gly Val Glu Gly Ser Thr Ser Ser Pr - #o Thr Ser Val Met Val                  345      - #           350      - #           355                  - - Gly Leu Gly Leu Ala Thr Val Val Thr Leu Th - #r Leu Ala Thr Ile Val              360          - #       365          - #       370                      - - Leu Gly Val Pro Arg Arg Arg Arg Ala Ala Al - #a His Leu Val Cys Pro          375              - #   380              - #   385                          - - Val Ser Ala Ser Gln                                                      390                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:7:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1338 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Oryctolagus - #cuniculus                                        (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 17..1261                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                               - - GAATTCGCGG CCGGCC TAC GGG CTC TTC GTT TGC CTA - #CTG CTC TGG GGA             49                                                                                          - #Tyr Gly Leu Phe Val Cys Leu Leu Leu Trp G - #ly                            - #  1               5 - #                 10                - - GGC TCG GAG CTG TGC TGC CCC CAG CCG CTC TG - #G TTC TGG CAG GGC GGG           97                                                                       Gly Ser Glu Leu Cys Cys Pro Gln Pro Leu Tr - #p Phe Trp Gln Gly Gly                        15     - #             20     - #             25                  - - ACC CGC CAG CCC GCG CCC TCC GTG ACG CCC GT - #G GTG GTG GAG TGT CTG          145                                                                       Thr Arg Gln Pro Ala Pro Ser Val Thr Pro Va - #l Val Val Glu Cys Leu                    30         - #         35         - #         40                      - - GAG GCC CGG CTC GTG GTC ACG GTC AGC AGG GA - #C CTT TTT GGC ACC GGG          193                                                                       Glu Ala Arg Leu Val Val Thr Val Ser Arg As - #p Leu Phe Gly Thr Gly                45             - #     50             - #     55                          - - AAG CTC ATC CAG GAG GCC GAC CTC AGC CTG GG - #C CCC GAG GGC TGC GAG          241                                                                       Lys Leu Ile Gln Glu Ala Asp Leu Ser Leu Gl - #y Pro Glu Gly Cys Glu            60                 - # 65                 - # 70                 - # 75       - - CCC CAG GCC TCC ACG GAC GCC GTG GTC AGG TT - #C GAG GTC GGG CTG CAT          289                                                                       Pro Gln Ala Ser Thr Asp Ala Val Val Arg Ph - #e Glu Val Gly Leu His                            80 - #                 85 - #                 90              - - GAA TGT GGT AAC AGC GTG CAG GTG ACT GAC GA - #C TCC CTG GTG TAC AGC          337                                                                       Glu Cys Gly Asn Ser Val Gln Val Thr Asp As - #p Ser Leu Val Tyr Ser                        95     - #            100     - #            105                  - - TCC TTC CTG CTC CAC GAC CCC CGC CCC GCG GG - #A AAC CTG TCC ATC CTC          385                                                                       Ser Phe Leu Leu His Asp Pro Arg Pro Ala Gl - #y Asn Leu Ser Ile Leu                   110          - #       115          - #       120                      - - AGG ACC AAC CGC GCC GAG GTC CCC ATC GAG TG - #C CGC TAC CCC AGG CAG          433                                                                       Arg Thr Asn Arg Ala Glu Val Pro Ile Glu Cy - #s Arg Tyr Pro Arg Gln               125              - #   130              - #   135                          - - GGC AAC GTG AGC AGC CGG GCG ATC CTG CCG AC - #C TGG GTG CCC TTC TGG          481                                                                       Gly Asn Val Ser Ser Arg Ala Ile Leu Pro Th - #r Trp Val Pro Phe Trp           140                 1 - #45                 1 - #50                 1 -      #55                                                                              - - ACC ACG GTA CTG TCA GAG GAG AGG CTG GTG TT - #C TCC CTG CGC CTC        ATG      529                                                                    Thr Thr Val Leu Ser Glu Glu Arg Leu Val Ph - #e Ser Leu Arg Leu Met                          160  - #               165  - #               170              - - GAG GAG AAC TGG AGC CGA GAA AAG ATG TCC CC - #C ACC TTC CAC CTG GGC          577                                                                       Glu Glu Asn Trp Ser Arg Glu Lys Met Ser Pr - #o Thr Phe His Leu Gly                       175      - #           180      - #           185                  - - GAC ACG GCC CAC CTG CAG GCA GAG GTC CGC AC - #G GGC AGC CAC CCG CCC          625                                                                       Asp Thr Ala His Leu Gln Ala Glu Val Arg Th - #r Gly Ser His Pro Pro                   190          - #       195          - #       200                      - - CTG CTG CTG TTC GTG GAT CGC TGC GTG GCC AC - #C CCG ACA CGG GAC CAG          673                                                                       Leu Leu Leu Phe Val Asp Arg Cys Val Ala Th - #r Pro Thr Arg Asp Gln               205              - #   210              - #   215                          - - AGC GGC TCC CCC TAT CAC ACC ATC GTG GAC TT - #G CAC GGC TGT CTT GTG          721                                                                       Ser Gly Ser Pro Tyr His Thr Ile Val Asp Le - #u His Gly Cys Leu Val           220                 2 - #25                 2 - #30                 2 -      #35                                                                              - - GAT GGC CTC TCC GAT GGG GCT TCC AAG TTC AA - #A GCC CCC AGG CCG        AAG      769                                                                    Asp Gly Leu Ser Asp Gly Ala Ser Lys Phe Ly - #s Ala Pro Arg Pro Lys                          240  - #               245  - #               250              - - CCG GAC GTG CTC CAG TTC ATG GTG GCC GTG TT - #C CAC TTC GCT AAT GAC          817                                                                       Pro Asp Val Leu Gln Phe Met Val Ala Val Ph - #e His Phe Ala Asn Asp                       255      - #           260      - #           265                  - - TCC AGG CAC ACG GTC TAC ATC ACG TGT CAC CT - #G AGG GTC ATT CCT GCC          865                                                                       Ser Arg His Thr Val Tyr Ile Thr Cys His Le - #u Arg Val Ile Pro Ala                   270          - #       275          - #       280                      - - CAG CAA GCC CCG GAC CGG CTC AAC AAG GCT TG - #T TCT TTC AAC CAG TCC          913                                                                       Gln Gln Ala Pro Asp Arg Leu Asn Lys Ala Cy - #s Ser Phe Asn Gln Ser               285              - #   290              - #   295                          - - TCC AGC AGC TGG GCC CCG GTG GAA GGC AGT GC - #A GAC ATC TGT GAG TGT          961                                                                       Ser Ser Ser Trp Ala Pro Val Glu Gly Ser Al - #a Asp Ile Cys Glu Cys           300                 3 - #05                 3 - #10                 3 -      #15                                                                              - - TGC GGC AAC GGT GAC TGT GAC CTC ATC GCA GG - #C TCC CCC ATG AAC        CAG     1009                                                                    Cys Gly Asn Gly Asp Cys Asp Leu Ile Ala Gl - #y Ser Pro Met Asn Gln                          320  - #               325  - #               330              - - AAC CAT GCT GCC CGG TCC TCT CTG CGA AGC CG - #C AGG CAC GTG ACG GAA         1057                                                                       Asn His Ala Ala Arg Ser Ser Leu Arg Ser Ar - #g Arg His Val Thr Glu                       335      - #           340      - #           345                  - - GAA GCA GAC GTC ACC GTG GGC CCG CTG ATC TT - #C CTG GGG AAG GCT GGT         1105                                                                       Glu Ala Asp Val Thr Val Gly Pro Leu Ile Ph - #e Leu Gly Lys Ala Gly                   350          - #       355          - #       360                      - - GAC CCT GCC GGC ACA GAG GGG CTG GCC TCT GC - #T GCG CAG GCG ACC CTG         1153                                                                       Asp Pro Ala Gly Thr Glu Gly Leu Ala Ser Al - #a Ala Gln Ala Thr Leu               365              - #   370              - #   375                          - - GTG CTG GGC CTT CGC ATG GCC ACC ATT GTG TT - #C CTG GCT GTG GCT GCT         1201                                                                       Val Leu Gly Leu Arg Met Ala Thr Ile Val Ph - #e Leu Ala Val Ala Ala           380                 3 - #85                 3 - #90                 3 -      #95                                                                              - - GTG GTC CTG GGC CTC ACC AGG GGG CGC CAC GC - #T GCT TCC CAC CCC        AGG     1249                                                                    Val Val Leu Gly Leu Thr Arg Gly Arg His Al - #a Ala Ser His Pro Arg                          400  - #               405  - #               410              - - TCT GCT TCC CAA TAAAAAATCA TGACTTCAAA AAAAAAAAAA AA - #AAAAAAAA             1301                                                                       Ser Ala Ser Gln                                                                           415                                                                - - AAAAAAAAAA AAAAAAAAAA AAAGCGGCCG CGAATTC      - #                       - #    1338                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:8:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 415 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                               - - Tyr Gly Leu Phe Val Cys Leu Leu Leu Trp Gl - #y Gly Ser Glu Leu        Cys                                                                               1               5 - #                 10 - #                 15             - - Cys Pro Gln Pro Leu Trp Phe Trp Gln Gly Gl - #y Thr Arg Gln Pro Ala                   20     - #             25     - #             30                  - - Pro Ser Val Thr Pro Val Val Val Glu Cys Le - #u Glu Ala Arg Leu Val               35         - #         40         - #         45                      - - Val Thr Val Ser Arg Asp Leu Phe Gly Thr Gl - #y Lys Leu Ile Gln Glu           50             - #     55             - #     60                          - - Ala Asp Leu Ser Leu Gly Pro Glu Gly Cys Gl - #u Pro Gln Ala Ser Thr       65                 - # 70                 - # 75                 - # 80       - - Asp Ala Val Val Arg Phe Glu Val Gly Leu Hi - #s Glu Cys Gly Asn Ser                       85 - #                 90 - #                 95              - - Val Gln Val Thr Asp Asp Ser Leu Val Tyr Se - #r Ser Phe Leu Leu His                  100      - #           105      - #           110                  - - Asp Pro Arg Pro Ala Gly Asn Leu Ser Ile Le - #u Arg Thr Asn Arg Ala              115          - #       120          - #       125                      - - Glu Val Pro Ile Glu Cys Arg Tyr Pro Arg Gl - #n Gly Asn Val Ser Ser          130              - #   135              - #   140                          - - Arg Ala Ile Leu Pro Thr Trp Val Pro Phe Tr - #p Thr Thr Val Leu Ser      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Glu Glu Arg Leu Val Phe Ser Leu Arg Leu Me - #t Glu Glu Asn Trp        Ser                                                                                             165  - #               170  - #               175             - - Arg Glu Lys Met Ser Pro Thr Phe His Leu Gl - #y Asp Thr Ala His Leu                  180      - #           185      - #           190                  - - Gln Ala Glu Val Arg Thr Gly Ser His Pro Pr - #o Leu Leu Leu Phe Val              195          - #       200          - #       205                      - - Asp Arg Cys Val Ala Thr Pro Thr Arg Asp Gl - #n Ser Gly Ser Pro Tyr          210              - #   215              - #   220                          - - His Thr Ile Val Asp Leu His Gly Cys Leu Va - #l Asp Gly Leu Ser Asp      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Gly Ala Ser Lys Phe Lys Ala Pro Arg Pro Ly - #s Pro Asp Val Leu        Gln                                                                                             245  - #               250  - #               255             - - Phe Met Val Ala Val Phe His Phe Ala Asn As - #p Ser Arg His Thr Val                  260      - #           265      - #           270                  - - Tyr Ile Thr Cys His Leu Arg Val Ile Pro Al - #a Gln Gln Ala Pro Asp              275          - #       280          - #       285                      - - Arg Leu Asn Lys Ala Cys Ser Phe Asn Gln Se - #r Ser Ser Ser Trp Ala          290              - #   295              - #   300                          - - Pro Val Glu Gly Ser Ala Asp Ile Cys Glu Cy - #s Cys Gly Asn Gly Asp      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Cys Asp Leu Ile Ala Gly Ser Pro Met Asn Gl - #n Asn His Ala Ala        Arg                                                                                             325  - #               330  - #               335             - - Ser Ser Leu Arg Ser Arg Arg His Val Thr Gl - #u Glu Ala Asp Val Thr                  340      - #           345      - #           350                  - - Val Gly Pro Leu Ile Phe Leu Gly Lys Ala Gl - #y Asp Pro Ala Gly Thr              355          - #       360          - #       365                      - - Glu Gly Leu Ala Ser Ala Ala Gln Ala Thr Le - #u Val Leu Gly Leu Arg          370              - #   375              - #   380                          - - Met Ala Thr Ile Val Phe Leu Ala Val Ala Al - #a Val Val Leu Gly Leu      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Thr Arg Gly Arg His Ala Ala Ser His Pro Ar - #g Ser Ala Ser Gln                         405  - #               410  - #               415              - -  - - (2) INFORMATION FOR SEQ ID NO:9:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2381 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Canis fam - #iliaris                                            (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 206..2353                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                               - - GAATTCCGGG AGCCCTGAAG GAAGCCGCAA GAACCCTGCC CGCACCTCCG CG -             #ACCTCAAG     60                                                                 - - ATGTCCACTC CACTGGAAGA CGGAGAATAC TGGATTGACC CCAACCAAGG AT -            #GCAACCTG    120                                                                 - - ATGCCATCAA GGTTTTCTGC AACATGGAGA CAGGTGAGAC CTGCGTATAC CC -            #ACCTACCT    180                                                                 - - GGCTGATTTG GTGGTACGTT TGGCC ATG GCA TGC AAA CAG - #AAA GGA GAC AGT          232                                                                                         - #          Met Ala Cys Lys Gln - # Lys Gly Asp Ser                          - #            1      - #         5                          - - GGG AGT CCC TCA AGC AGG TTT AGT GCA GAT TG - #G AGC ACC TAC AGG TCA          280                                                                       Gly Ser Pro Ser Ser Arg Phe Ser Ala Asp Tr - #p Ser Thr Tyr Arg Ser            10                 - # 15                 - # 20                 - # 25       - - CTT TCT TTA TTC TTC ATC CTT GTG ACT TCA GT - #G AAC TCA GTA GGT GTT          328                                                                       Leu Ser Leu Phe Phe Ile Leu Val Thr Ser Va - #l Asn Ser Val Gly Val                            30 - #                 35 - #                 40              - - ATG CAG TTG GTG AAT CCC ATC TTC CCA GGT AC - #T GTC ATT TGC CAT GAA          376                                                                       Met Gln Leu Val Asn Pro Ile Phe Pro Gly Th - #r Val Ile Cys His Glu                        45     - #             50     - #             55                  - - AAT AAA ATG ACA GTG GAA TTT CCA AGG GAT CT - #T GGC ACC AAA AAA TGG          424                                                                       Asn Lys Met Thr Val Glu Phe Pro Arg Asp Le - #u Gly Thr Lys Lys Trp                    60         - #         65         - #         70                      - - CAT GCA TCT GTG GTG GAT CCA TTT AGT TTT GA - #A TTG TTG AAC TGT ACT          472                                                                       His Ala Ser Val Val Asp Pro Phe Ser Phe Gl - #u Leu Leu Asn Cys Thr                75             - #     80             - #     85                          - - TCT ATC CTG GAC CCA GAA AAG CTC ACC CTG AA - #G GCC CCA TAT GAG ACC          520                                                                       Ser Ile Leu Asp Pro Glu Lys Leu Thr Leu Ly - #s Ala Pro Tyr Glu Thr            90                 - # 95                 - #100                 - #105       - - TGT AGC AGG AGA GTG CTT GGC CAG CAT CAG AT - #G GCC ATC AGA CTC ACG          568                                                                       Cys Ser Arg Arg Val Leu Gly Gln His Gln Me - #t Ala Ile Arg Leu Thr                           110  - #               115  - #               120              - - GAC AAC AAT GCT GCT TCA AGA CAT AAG GCT TT - #C ATG TAT CAG ATC AGC          616                                                                       Asp Asn Asn Ala Ala Ser Arg His Lys Ala Ph - #e Met Tyr Gln Ile Ser                       125      - #           130      - #           135                  - - TGT CCA GTT ATG CAA ACA GAA GAA ACC CAT GA - #G CAT GCA GGA TCC ACA          664                                                                       Cys Pro Val Met Gln Thr Glu Glu Thr His Gl - #u His Ala Gly Ser Thr                   140          - #       145          - #       150                      - - ATC TGC ACA AAA GAT TCC ATG TCT TTT ACC TT - #T AAC ATT ATT CCT GGC          712                                                                       Ile Cys Thr Lys Asp Ser Met Ser Phe Thr Ph - #e Asn Ile Ile Pro Gly               155              - #   160              - #   165                          - - ATG GCT GAT GAA AAT ACG AAT CCC AGT GGT GG - #G AAA TGG ATG ATG GAG          760                                                                       Met Ala Asp Glu Asn Thr Asn Pro Ser Gly Gl - #y Lys Trp Met Met Glu           170                 1 - #75                 1 - #80                 1 -      #85                                                                              - - GTT GAT GAT GCA AAA GCT CAA AAT CTG ACT CT - #T CGG GAG GCC TTG        ATG      808                                                                    Val Asp Asp Ala Lys Ala Gln Asn Leu Thr Le - #u Arg Glu Ala Leu Met                          190  - #               195  - #               200              - - CAA GGA TAT AAT TTC CTG TTT GAT AGC CAC AG - #G CTC AGT GTC CAA GTG          856                                                                       Gln Gly Tyr Asn Phe Leu Phe Asp Ser His Ar - #g Leu Ser Val Gln Val                       205      - #           210      - #           215                  - - TCA TTC AAT GCC ACT GGA GTC ACT CAC TAC AT - #G CAA GGT AAC AGT CAC          904                                                                       Ser Phe Asn Ala Thr Gly Val Thr His Tyr Me - #t Gln Gly Asn Ser His                   220          - #       225          - #       230                      - - CTC TAC ACA GTG CCT CTG AAG CTT ATA CAC AC - #A TCT CCT GGG CAG AAG          952                                                                       Leu Tyr Thr Val Pro Leu Lys Leu Ile His Th - #r Ser Pro Gly Gln Lys               235              - #   240              - #   245                          - - ATC ATC TTA ACA ACA CGA GTA CTT TGT ATG TC - #A GAT CCC GTG ACC TGT         1000                                                                       Ile Ile Leu Thr Thr Arg Val Leu Cys Met Se - #r Asp Pro Val Thr Cys           250                 2 - #55                 2 - #60                 2 -      #65                                                                              - - AAC GCC ACA CAC ATG ACC CTC ACC ATA CCA GA - #G TTT CCT GGG AAA        CTA     1048                                                                    Asn Ala Thr His Met Thr Leu Thr Ile Pro Gl - #u Phe Pro Gly Lys Leu                          270  - #               275  - #               280              - - CAG TCT GTG AGA TTT GAA AAC ACG AAC TTT CG - #T GTA AGC CAG CTG CAC         1096                                                                       Gln Ser Val Arg Phe Glu Asn Thr Asn Phe Ar - #g Val Ser Gln Leu His                       285      - #           290      - #           295                  - - AAC CAT GGG ATT GAT AAA GAA GAA TTA AAC GG - #C TTG AGG TTA CAC TTC         1144                                                                       Asn His Gly Ile Asp Lys Glu Glu Leu Asn Gl - #y Leu Arg Leu His Phe                   300          - #       305          - #       310                      - - AGC AAA TCT CTT CTC AAA ATG AAC TCC TCT GA - #A AAA TGC CTA CTC TAT         1192                                                                       Ser Lys Ser Leu Leu Lys Met Asn Ser Ser Gl - #u Lys Cys Leu Leu Tyr               315              - #   320              - #   325                          - - CAG TTC TAC TTA GCA TCT CTC AAG CTG ACC TT - #T GCC TTT GAA CGG GAC         1240                                                                       Gln Phe Tyr Leu Ala Ser Leu Lys Leu Thr Ph - #e Ala Phe Glu Arg Asp           330                 3 - #35                 3 - #40                 3 -      #45                                                                              - - ACG GTT TCC ACA GTG GTT TAT CCT GAG TGT GT - #T TGT GAG CCA CCA        GTT     1288                                                                    Thr Val Ser Thr Val Val Tyr Pro Glu Cys Va - #l Cys Glu Pro Pro Val                          350  - #               355  - #               360              - - ACT ATA GTT ACA GGT GAC CTG TGT ACC CAG GA - #T GGG TTT ATG GAT GTC         1336                                                                       Thr Ile Val Thr Gly Asp Leu Cys Thr Gln As - #p Gly Phe Met Asp Val                       365      - #           370      - #           375                  - - AAG GTC TAC AGC CAC CAA ACA AAA CCA GCT CT - #A AAC TTG GAT ACC CTC         1384                                                                       Lys Val Tyr Ser His Gln Thr Lys Pro Ala Le - #u Asn Leu Asp Thr Leu                   380          - #       385          - #       390                      - - AGA GTG GGA GAC TCC TCC TGC CAA CCT ACT TT - #C AAG GCT CCA TCA CAA         1432                                                                       Arg Val Gly Asp Ser Ser Cys Gln Pro Thr Ph - #e Lys Ala Pro Ser Gln               395              - #   400              - #   405                          - - GGG TTG ACA CTG TTT CAC ATC CCC CTA AAT GG - #A TGT GGA ACA AGA CTT         1480                                                                       Gly Leu Thr Leu Phe His Ile Pro Leu Asn Gl - #y Cys Gly Thr Arg Leu           410                 4 - #15                 4 - #20                 4 -      #25                                                                              - - AAG TTC AAA GGT GAC ACA GTC ATC TAT GAA AA - #T GAA ATA CAT GCT        CTC     1528                                                                    Lys Phe Lys Gly Asp Thr Val Ile Tyr Glu As - #n Glu Ile His Ala Leu                          430  - #               435  - #               440              - - TGG ACA GAT CTC CCT CCA AGC ACA ATT TCC AG - #A GAT AGT GAA TTC AGA         1576                                                                       Trp Thr Asp Leu Pro Pro Ser Thr Ile Ser Ar - #g Asp Ser Glu Phe Arg                       445      - #           450      - #           455                  - - ATG ACT GTG AAG TGC CAT TAC AGC AGA GAT GA - #C CTG CTG ATA AAT ACC         1624                                                                       Met Thr Val Lys Cys His Tyr Ser Arg Asp As - #p Leu Leu Ile Asn Thr                   460          - #       465          - #       470                      - - AAT GTC CAA AGT CTT CCT CCT CCC GTG GCC TC - #A GTG AGG CCT GGT CCA         1672                                                                       Asn Val Gln Ser Leu Pro Pro Pro Val Ala Se - #r Val Arg Pro Gly Pro               475              - #   480              - #   485                          - - CTT GCC TTA ATC CTG CAA ACC TAC CCA GAT AA - #A TCC TAT TTG CGA CCC         1720                                                                       Leu Ala Leu Ile Leu Gln Thr Tyr Pro Asp Ly - #s Ser Tyr Leu Arg Pro           490                 4 - #95                 5 - #00                 5 -      #05                                                                              - - TAT GGG GAT AAG GAG TAT CCT GTG GTG AGA TA - #C CTC CGC CAA CCA        ATT     1768                                                                    Tyr Gly Asp Lys Glu Tyr Pro Val Val Arg Ty - #r Leu Arg Gln Pro Ile                          510  - #               515  - #               520              - - TAC CTG GAA GTG AAA GTC CTA AAT AGG GCT GA - #C CCC AAC ATC AAG CTG         1816                                                                       Tyr Leu Glu Val Lys Val Leu Asn Arg Ala As - #p Pro Asn Ile Lys Leu                       525      - #           530      - #           535                  - - GTC TTA GAT GAT TGC TGG GCA ACA CCC ACC AT - #G GAC CCA GCC TCA CTC         1864                                                                       Val Leu Asp Asp Cys Trp Ala Thr Pro Thr Me - #t Asp Pro Ala Ser Leu                   540          - #       545          - #       550                      - - CCC CAG TGG AAT ATT GTC ATG GAT GGC TGT GA - #A TAC AAT CTG GAC AAC         1912                                                                       Pro Gln Trp Asn Ile Val Met Asp Gly Cys Gl - #u Tyr Asn Leu Asp Asn               555              - #   560              - #   565                          - - TAC AGA ACG ACC TTC CAT CCA GTT GGC TCC TC - #T GTG ACC TAC CCT ACT         1960                                                                       Tyr Arg Thr Thr Phe His Pro Val Gly Ser Se - #r Val Thr Tyr Pro Thr           570                 5 - #75                 5 - #80                 5 -      #85                                                                              - - CAC TAT CAG AGG TTT GAT GTG AAG ACC TTT GC - #C TTT ATA TCA GAG        GCC     2008                                                                    His Tyr Gln Arg Phe Asp Val Lys Thr Phe Al - #a Phe Ile Ser Glu Ala                          590  - #               595  - #               600              - - CAA GTG CTT TCT AGC CTG GTC TAC TTC CAC TG - #C ACC GCA TTA ATC TGC         2056                                                                       Gln Val Leu Ser Ser Leu Val Tyr Phe His Cy - #s Thr Ala Leu Ile Cys                       605      - #           610      - #           615                  - - AAT CGA CTG TCT CCT GAC TCC CCT CTG TGT TC - #T GTG ACT TGC CCT GTA         2104                                                                       Asn Arg Leu Ser Pro Asp Ser Pro Leu Cys Se - #r Val Thr Cys Pro Val                   620          - #       625          - #       630                      - - TCA TCC AGG CAC AGG CGA GCC ACA GGC AGT AC - #T GAA GAA GAG AAG ATG         2152                                                                       Ser Ser Arg His Arg Arg Ala Thr Gly Ser Th - #r Glu Glu Glu Lys Met               635              - #   640              - #   645                          - - ATA GTA AGT CTC CCG GGA CCC ATC CTC CTG TT - #G GCA GAC AGC TCT TCA         2200                                                                       Ile Val Ser Leu Pro Gly Pro Ile Leu Leu Le - #u Ala Asp Ser Ser Ser           650                 6 - #55                 6 - #60                 6 -      #65                                                                              - - CTC AGA GAT GGT GTG GAC TCA AAA GGG CAC AG - #G GCT GCT GGA TAT        GTT     2248                                                                    Leu Arg Asp Gly Val Asp Ser Lys Gly His Ar - #g Ala Ala Gly Tyr Val                          670  - #               675  - #               680              - - GCT TTT AAA ACT GTA GTG GCT GTG GCT GCC TT - #A GCA GGC CTT GTG GCT         2296                                                                       Ala Phe Lys Thr Val Val Ala Val Ala Ala Le - #u Ala Gly Leu Val Ala                       685      - #           690      - #           695                  - - GCT CTA GGT CTC ATC ATC TAC CTG CGT AAG AA - #A AGA ACC ATG GTG TTA         2344                                                                       Ala Leu Gly Leu Ile Ile Tyr Leu Arg Lys Ly - #s Arg Thr Met Val Leu                   700          - #       705          - #       710                      - - AAT CAC TAAGGATTTT CAAATAAAGT GTCCGGAATT C    - #                       - #    2381                                                                     Asn His                                                                           715                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:10:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 715 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                              - - Met Ala Cys Lys Gln Lys Gly Asp Ser Gly Se - #r Pro Ser Ser Arg        Phe                                                                               1               5 - #                 10 - #                 15             - - Ser Ala Asp Trp Ser Thr Tyr Arg Ser Leu Se - #r Leu Phe Phe Ile Leu                   20     - #             25     - #             30                  - - Val Thr Ser Val Asn Ser Val Gly Val Met Gl - #n Leu Val Asn Pro Ile               35         - #         40         - #         45                      - - Phe Pro Gly Thr Val Ile Cys His Glu Asn Ly - #s Met Thr Val Glu Phe           50             - #     55             - #     60                          - - Pro Arg Asp Leu Gly Thr Lys Lys Trp His Al - #a Ser Val Val Asp Pro       65                 - # 70                 - # 75                 - # 80       - - Phe Ser Phe Glu Leu Leu Asn Cys Thr Ser Il - #e Leu Asp Pro Glu Lys                       85 - #                 90 - #                 95              - - Leu Thr Leu Lys Ala Pro Tyr Glu Thr Cys Se - #r Arg Arg Val Leu Gly                  100      - #           105      - #           110                  - - Gln His Gln Met Ala Ile Arg Leu Thr Asp As - #n Asn Ala Ala Ser Arg              115          - #       120          - #       125                      - - His Lys Ala Phe Met Tyr Gln Ile Ser Cys Pr - #o Val Met Gln Thr Glu          130              - #   135              - #   140                          - - Glu Thr His Glu His Ala Gly Ser Thr Ile Cy - #s Thr Lys Asp Ser Met      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ser Phe Thr Phe Asn Ile Ile Pro Gly Met Al - #a Asp Glu Asn Thr        Asn                                                                                             165  - #               170  - #               175             - - Pro Ser Gly Gly Lys Trp Met Met Glu Val As - #p Asp Ala Lys Ala Gln                  180      - #           185      - #           190                  - - Asn Leu Thr Leu Arg Glu Ala Leu Met Gln Gl - #y Tyr Asn Phe Leu Phe              195          - #       200          - #       205                      - - Asp Ser His Arg Leu Ser Val Gln Val Ser Ph - #e Asn Ala Thr Gly Val          210              - #   215              - #   220                          - - Thr His Tyr Met Gln Gly Asn Ser His Leu Ty - #r Thr Val Pro Leu Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Leu Ile His Thr Ser Pro Gly Gln Lys Ile Il - #e Leu Thr Thr Arg        Val                                                                                             245  - #               250  - #               255             - - Leu Cys Met Ser Asp Pro Val Thr Cys Asn Al - #a Thr His Met Thr Leu                  260      - #           265      - #           270                  - - Thr Ile Pro Glu Phe Pro Gly Lys Leu Gln Se - #r Val Arg Phe Glu Asn              275          - #       280          - #       285                      - - Thr Asn Phe Arg Val Ser Gln Leu His Asn Hi - #s Gly Ile Asp Lys Glu          290              - #   295              - #   300                          - - Glu Leu Asn Gly Leu Arg Leu His Phe Ser Ly - #s Ser Leu Leu Lys Met      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Asn Ser Ser Glu Lys Cys Leu Leu Tyr Gln Ph - #e Tyr Leu Ala Ser        Leu                                                                                             325  - #               330  - #               335             - - Lys Leu Thr Phe Ala Phe Glu Arg Asp Thr Va - #l Ser Thr Val Val Tyr                  340      - #           345      - #           350                  - - Pro Glu Cys Val Cys Glu Pro Pro Val Thr Il - #e Val Thr Gly Asp Leu              355          - #       360          - #       365                      - - Cys Thr Gln Asp Gly Phe Met Asp Val Lys Va - #l Tyr Ser His Gln Thr          370              - #   375              - #   380                          - - Lys Pro Ala Leu Asn Leu Asp Thr Leu Arg Va - #l Gly Asp Ser Ser Cys      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Gln Pro Thr Phe Lys Ala Pro Ser Gln Gly Le - #u Thr Leu Phe His        Ile                                                                                             405  - #               410  - #               415             - - Pro Leu Asn Gly Cys Gly Thr Arg Leu Lys Ph - #e Lys Gly Asp Thr Val                  420      - #           425      - #           430                  - - Ile Tyr Glu Asn Glu Ile His Ala Leu Trp Th - #r Asp Leu Pro Pro Ser              435          - #       440          - #       445                      - - Thr Ile Ser Arg Asp Ser Glu Phe Arg Met Th - #r Val Lys Cys His Tyr          450              - #   455              - #   460                          - - Ser Arg Asp Asp Leu Leu Ile Asn Thr Asn Va - #l Gln Ser Leu Pro Pro      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Pro Val Ala Ser Val Arg Pro Gly Pro Leu Al - #a Leu Ile Leu Gln        Thr                                                                                             485  - #               490  - #               495             - - Tyr Pro Asp Lys Ser Tyr Leu Arg Pro Tyr Gl - #y Asp Lys Glu Tyr Pro                  500      - #           505      - #           510                  - - Val Val Arg Tyr Leu Arg Gln Pro Ile Tyr Le - #u Glu Val Lys Val Leu              515          - #       520          - #       525                      - - Asn Arg Ala Asp Pro Asn Ile Lys Leu Val Le - #u Asp Asp Cys Trp Ala          530              - #   535              - #   540                          - - Thr Pro Thr Met Asp Pro Ala Ser Leu Pro Gl - #n Trp Asn Ile Val Met      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Asp Gly Cys Glu Tyr Asn Leu Asp Asn Tyr Ar - #g Thr Thr Phe His        Pro                                                                                             565  - #               570  - #               575             - - Val Gly Ser Ser Val Thr Tyr Pro Thr His Ty - #r Gln Arg Phe Asp Val                  580      - #           585      - #           590                  - - Lys Thr Phe Ala Phe Ile Ser Glu Ala Gln Va - #l Leu Ser Ser Leu Val              595          - #       600          - #       605                      - - Tyr Phe His Cys Thr Ala Leu Ile Cys Asn Ar - #g Leu Ser Pro Asp Ser          610              - #   615              - #   620                          - - Pro Leu Cys Ser Val Thr Cys Pro Val Ser Se - #r Arg His Arg Arg Ala      625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - Thr Gly Ser Thr Glu Glu Glu Lys Met Ile Va - #l Ser Leu Pro Gly        Pro                                                                                             645  - #               650  - #               655             - - Ile Leu Leu Leu Ala Asp Ser Ser Ser Leu Ar - #g Asp Gly Val Asp Ser                  660      - #           665      - #           670                  - - Lys Gly His Arg Ala Ala Gly Tyr Val Ala Ph - #e Lys Thr Val Val Ala              675          - #       680          - #       685                      - - Val Ala Ala Leu Ala Gly Leu Val Ala Ala Le - #u Gly Leu Ile Ile Tyr          690              - #   695              - #   700                          - - Leu Arg Lys Lys Arg Thr Met Val Leu Asn Hi - #s                          705                 7 - #10                 7 - #15                            - -  - - (2) INFORMATION FOR SEQ ID NO:11:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1325 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Canis fam - #iliaris                                            (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 13..1293                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                              - - GAATTCCGGG CT ATG GGG CTG AGC TAT GGA ATT TTC - # ATC TGT TTT CTG             48                                                                                     Met Gly Le - #u Ser Tyr Gly Ile Phe Ile Cys Phe Leu                             1  - #             5     - #             10                    - - CTC CTG GGA GGC ATG GAG CTG TGC TGC CCC CA - #G ACC ATC TGG CCA ACT           96                                                                       Leu Leu Gly Gly Met Glu Leu Cys Cys Pro Gl - #n Thr Ile Trp Pro Thr                    15         - #         20         - #         25                      - - GAG ACC TAC TAC CCA TTG ACA TCT AGG CCC CC - #A GTA ATG GTG GAC TGT          144                                                                       Glu Thr Tyr Tyr Pro Leu Thr Ser Arg Pro Pr - #o Val Met Val Asp Cys                30             - #     35             - #     40                          - - CTG GAG TCC CAG CTG GTG GTC ACT GTC AGC AA - #A GAC CTT TTT GGT ACT          192                                                                       Leu Glu Ser Gln Leu Val Val Thr Val Ser Ly - #s Asp Leu Phe Gly Thr            45                 - # 50                 - # 55                 - # 60       - - GGG AAG CTC ATC AGG CCA GCA GAC CTC ACC CT - #G GGT CCA GAG AAC TGT          240                                                                       Gly Lys Leu Ile Arg Pro Ala Asp Leu Thr Le - #u Gly Pro Glu Asn Cys                            65 - #                 70 - #                 75              - - GAG CCC CTG GTC TCC ATG GAC ACG GAT GAT GT - #G GTC AGG TTT GAG GTT          288                                                                       Glu Pro Leu Val Ser Met Asp Thr Asp Asp Va - #l Val Arg Phe Glu Val                        80     - #             85     - #             90                  - - GGG CTG CAC GAG TGT GGC AGC AGG GTG CAG GT - #G ACT GAC AAT GCT CTG          336                                                                       Gly Leu His Glu Cys Gly Ser Arg Val Gln Va - #l Thr Asp Asn Ala Leu                    95         - #        100         - #        105                      - - GTG TAC AGC ACC TTC CTG ATC CAC AGC CCC CG - #C CCT GCG GGC AAC CTG          384                                                                       Val Tyr Ser Thr Phe Leu Ile His Ser Pro Ar - #g Pro Ala Gly Asn Leu               110              - #   115              - #   120                          - - TCC ATC CTG AGA ACT AAT CGT GCC GAG GTT CC - #C ATC GAG TGC CAC TAC          432                                                                       Ser Ile Leu Arg Thr Asn Arg Ala Glu Val Pr - #o Ile Glu Cys His Tyr           125                 1 - #30                 1 - #35                 1 -      #40                                                                              - - CCC AGG CAC AGC AAT GTG AGC AGC CAG GCC AT - #C CTG CCC ACT TGG        GTG      480                                                                    Pro Arg His Ser Asn Val Ser Ser Gln Ala Il - #e Leu Pro Thr Trp Val                          145  - #               150  - #               155              - - CCC TTC AGG ACC ACA ATG CTC TTC GAG GAG AA - #G CTA GTT TTC TCT CTC          528                                                                       Pro Phe Arg Thr Thr Met Leu Phe Glu Glu Ly - #s Leu Val Phe Ser Leu                       160      - #           165      - #           170                  - - CGC CTA ATG GAG GAG GAC TGG GGC TCC GAG AA - #G CAA TCC CCC ACA TTC          576                                                                       Arg Leu Met Glu Glu Asp Trp Gly Ser Glu Ly - #s Gln Ser Pro Thr Phe                   175          - #       180          - #       185                      - - CAG CTG GGA GAC ATA GCC CAC CTC CAG GCT GA - #A GTC CAC ACT GGC AGC          624                                                                       Gln Leu Gly Asp Ile Ala His Leu Gln Ala Gl - #u Val His Thr Gly Ser               190              - #   195              - #   200                          - - CAT ATG CCA CTG CGA CTT TTT GTG GAC CAC TG - #T GTG GCC ACG CTG ACA          672                                                                       His Met Pro Leu Arg Leu Phe Val Asp His Cy - #s Val Ala Thr Leu Thr           205                 2 - #10                 2 - #15                 2 -      #20                                                                              - - CCA GAT CGG AAT GCC TTC CTT CAT CAC AAA AT - #T GTG GAC TTC CAT        GGC      720                                                                    Pro Asp Arg Asn Ala Phe Leu His His Lys Il - #e Val Asp Phe His Gly                          225  - #               230  - #               235              - - TGT CTT GTG GAT GGT CTC TAC AAT TCC TCT TC - #A GCC TTC AAA GCC CCC          768                                                                       Cys Leu Val Asp Gly Leu Tyr Asn Ser Ser Se - #r Ala Phe Lys Ala Pro                       240      - #           245      - #           250                  - - AGA CCC AGG CCA GAG ACT CTT CAG TTC ACA GT - #G GAT GTT TTC CAC TTT          816                                                                       Arg Pro Arg Pro Glu Thr Leu Gln Phe Thr Va - #l Asp Val Phe His Phe                   255          - #       260          - #       265                      - - GCT AAG GAC TCA AGA AAC ACG ATC TAT ATC AC - #C TGC CAT CTG AAG GTC          864                                                                       Ala Lys Asp Ser Arg Asn Thr Ile Tyr Ile Th - #r Cys His Leu Lys Val               270              - #   275              - #   280                          - - ACT CCG GCT GAC CGA GTC CCA GAC CAG CTA AA - #C AAA GCT TGT TCC TTC          912                                                                       Thr Pro Ala Asp Arg Val Pro Asp Gln Leu As - #n Lys Ala Cys Ser Phe           285                 2 - #90                 2 - #95                 3 -      #00                                                                              - - ATC AAG TCT ACC AAG AGG TGG TAC CCT GTA GA - #A GGC TCG GCT GAT        ATT      960                                                                    Ile Lys Ser Thr Lys Arg Trp Tyr Pro Val Gl - #u Gly Ser Ala Asp Ile                          305  - #               310  - #               315              - - TGT CGC TGT TGT AAC AAA GGC AGC TGT GGC CT - #T CCA GGC CGG TCC AGG         1008                                                                       Cys Arg Cys Cys Asn Lys Gly Ser Cys Gly Le - #u Pro Gly Arg Ser Arg                       320      - #           325      - #           330                  - - AGG CTG TCC CAC CTA GAG AGA GGG TGG CGC AA - #G TCT GTT TCC CAC ACT         1056                                                                       Arg Leu Ser His Leu Glu Arg Gly Trp Arg Ly - #s Ser Val Ser His Thr                   335          - #       340          - #       345                      - - AGA AAT CGC AGG CAC GTG ACT GAA GAA GCA GA - #G ATC ACC GTG GGG CCT         1104                                                                       Arg Asn Arg Arg His Val Thr Glu Glu Ala Gl - #u Ile Thr Val Gly Pro               350              - #   355              - #   360                          - - CTG ATC TTC CTG GGA AAG GCT AGT GAT CAT GG - #T ATA GAG GGG TCA ACC         1152                                                                       Leu Ile Phe Leu Gly Lys Ala Ser Asp His Gl - #y Ile Glu Gly Ser Thr           365                 3 - #70                 3 - #75                 3 -      #80                                                                              - - TCT CCT CAC ACC TCT GTG ATG TTG GGC TTA GG - #C CTG GCC ACG GTG        GTA     1200                                                                    Ser Pro His Thr Ser Val Met Leu Gly Leu Gl - #y Leu Ala Thr Val Val                          385  - #               390  - #               395              - - TCC CTG ACT CTA GCT ACC ATT GTC CTG GTC CT - #T GCC AAG AGG CAT CGT         1248                                                                       Ser Leu Thr Leu Ala Thr Ile Val Leu Val Le - #u Ala Lys Arg His Arg                       400      - #           405      - #           410                  - - ACT GCT TCC CAC CCT GTG ATA TGC CCT GCA TC - #T GTC TCC CAA             TAAAAGAATA  1300                                                                Thr Ala Ser His Pro Val Ile Cys Pro Ala Se - #r Val Ser Gln                           415          - #       420          - #       425                      - - AGCAAAAAAA AAAAAACCGG AATTC          - #                  - #                 1325                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:12:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 426 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                              - - Met Gly Leu Ser Tyr Gly Ile Phe Ile Cys Ph - #e Leu Leu Leu Gly Gly        1               5 - #                 10 - #                 15              - - Met Glu Leu Cys Cys Pro Gln Thr Ile Trp Pr - #o Thr Glu Thr Tyr Tyr                   20     - #             25     - #             30                  - - Pro Leu Thr Ser Arg Pro Pro Val Met Val As - #p Cys Leu Glu Ser Gln               35         - #         40         - #         45                      - - Leu Val Val Thr Val Ser Lys Asp Leu Phe Gl - #y Thr Gly Lys Leu Ile           50             - #     55             - #     60                          - - Arg Pro Ala Asp Leu Thr Leu Gly Pro Glu As - #n Cys Glu Pro Leu Val       65                 - # 70                 - # 75                 - # 80       - - Ser Met Asp Thr Asp Asp Val Val Arg Phe Gl - #u Val Gly Leu His Glu                       85 - #                 90 - #                 95              - - Cys Gly Ser Arg Val Gln Val Thr Asp Asn Al - #a Leu Val Tyr Ser Thr                  100      - #           105      - #           110                  - - Phe Leu Ile His Ser Pro Arg Pro Ala Gly As - #n Leu Ser Ile Leu Arg              115          - #       120          - #       125                      - - Thr Asn Arg Ala Glu Val Pro Ile Glu Cys Hi - #s Tyr Pro Arg His Ser          130              - #   135              - #   140                          - - Asn Val Ser Ser Gln Ala Ile Leu Pro Thr Tr - #p Val Pro Phe Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Thr Met Leu Phe Glu Glu Lys Leu Val Phe Se - #r Leu Arg Leu Met        Glu                                                                                             165  - #               170  - #               175             - - Glu Asp Trp Gly Ser Glu Lys Gln Ser Pro Th - #r Phe Gln Leu Gly Asp                  180      - #           185      - #           190                  - - Ile Ala His Leu Gln Ala Glu Val His Thr Gl - #y Ser His Met Pro Leu              195          - #       200          - #       205                      - - Arg Leu Phe Val Asp His Cys Val Ala Thr Le - #u Thr Pro Asp Arg Asn          210              - #   215              - #   220                          - - Ala Phe Leu His His Lys Ile Val Asp Phe Hi - #s Gly Cys Leu Val Asp      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Gly Leu Tyr Asn Ser Ser Ser Ala Phe Lys Al - #a Pro Arg Pro Arg        Pro                                                                                             245  - #               250  - #               255             - - Glu Thr Leu Gln Phe Thr Val Asp Val Phe Hi - #s Phe Ala Lys Asp Ser                  260      - #           265      - #           270                  - - Arg Asn Thr Ile Tyr Ile Thr Cys His Leu Ly - #s Val Thr Pro Ala Asp              275          - #       280          - #       285                      - - Arg Val Pro Asp Gln Leu Asn Lys Ala Cys Se - #r Phe Ile Lys Ser Thr          290              - #   295              - #   300                          - - Lys Arg Trp Tyr Pro Val Glu Gly Ser Ala As - #p Ile Cys Arg Cys Cys      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Asn Lys Gly Ser Cys Gly Leu Pro Gly Arg Se - #r Arg Arg Leu Ser        His                                                                                             325  - #               330  - #               335             - - Leu Glu Arg Gly Trp Arg Lys Ser Val Ser Hi - #s Thr Arg Asn Arg Arg                  340      - #           345      - #           350                  - - His Val Thr Glu Glu Ala Glu Ile Thr Val Gl - #y Pro Leu Ile Phe Leu              355          - #       360          - #       365                      - - Gly Lys Ala Ser Asp His Gly Ile Glu Gly Se - #r Thr Ser Pro His Thr          370              - #   375              - #   380                          - - Ser Val Met Leu Gly Leu Gly Leu Ala Thr Va - #l Val Ser Leu Thr Leu      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ala Thr Ile Val Leu Val Leu Ala Lys Arg Hi - #s Arg Thr Ala Ser        His                                                                                             405  - #               410  - #               415             - - Pro Val Ile Cys Pro Ala Ser Val Ser Gln                                              420      - #           425                                         - -  - - (2) INFORMATION FOR SEQ ID NO:13:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2236 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Felis dom - #esticus                                            (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 28..2175                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                              - - GAATTCGCGG CCGCGATACT TTTGGCT ATG GCC TCC AGA CAG - #AAA GGA GAT             51                                                                                          - #            Met Ala Ser Ar - #g Gln Lys Gly Asp                            - #              1    - #           5                        - - AGT GGG AGT CCT TCA AGC TGG TTT AAT GCA GA - #T TGG AGC ACC TAC AGG           99                                                                       Ser Gly Ser Pro Ser Ser Trp Phe Asn Ala As - #p Trp Ser Thr Tyr Arg                10             - #     15             - #     20                          - - TCA CTT TTT CTA CTC TTT ATC CTC GTG ACT TC - #A GTG AAT TCC ATA GGT          147                                                                       Ser Leu Phe Leu Leu Phe Ile Leu Val Thr Se - #r Val Asn Ser Ile Gly            25                 - # 30                 - # 35                 - # 40       - - GTT TTG CAG TTG GTG AAT CCT GTC TTC CCA GG - #T ACT GTC ACT TGC TAT          195                                                                       Val Leu Gln Leu Val Asn Pro Val Phe Pro Gl - #y Thr Val Thr Cys Tyr                            45 - #                 50 - #                 55              - - GAA ACT AGA ATG GCA GTG GAA TTT CCA AGT GA - #T TTT GGC ACC AAA AAA          243                                                                       Glu Thr Arg Met Ala Val Glu Phe Pro Ser As - #p Phe Gly Thr Lys Lys                        60     - #             65     - #             70                  - - TGG CAT ACA TCT GTG GTG GAT CCC TTT AGT TT - #T GAA TTG TTG AAC TGC          291                                                                       Trp His Thr Ser Val Val Asp Pro Phe Ser Ph - #e Glu Leu Leu Asn Cys                    75         - #         80         - #         85                      - - ACT TAC ATC TTG GAT CCA GAA AAT CTC ACC TT - #A AAG GCC CCA TAT GAG          339                                                                       Thr Tyr Ile Leu Asp Pro Glu Asn Leu Thr Le - #u Lys Ala Pro Tyr Glu                90             - #     95             - #    100                          - - ACC TGT ACC AGA AGA ACG CTT GGC CAG CAC CG - #G ATG ATC ATC AGA CTC          387                                                                       Thr Cys Thr Arg Arg Thr Leu Gly Gln His Ar - #g Met Ile Ile Arg Leu           105                 1 - #10                 1 - #15                 1 -      #20                                                                              - - AAG GAC CAC AAT GCT GCT TCA AGA CAT AAC AG - #T TTG ATG TAT CAG        ATC      435                                                                    Lys Asp His Asn Ala Ala Ser Arg His Asn Se - #r Leu Met Tyr Gln Ile                          125  - #               130  - #               135              - - AAC TGT CCA GTT ATG CAA GCA GAA GAA ACC CA - #T GAG CAT GCA GGA TCC          483                                                                       Asn Cys Pro Val Met Gln Ala Glu Glu Thr Hi - #s Glu His Ala Gly Ser                       140      - #           145      - #           150                  - - ACT ATC TGC ACA AAG GAT TCC ATG TCT TTT AC - #C TTT AAT GTC ATT CCT          531                                                                       Thr Ile Cys Thr Lys Asp Ser Met Ser Phe Th - #r Phe Asn Val Ile Pro                   155          - #       160          - #       165                      - - GGC CTG GCT GAT GAA AAT ACG GAT ATC AAG AA - #T CCG ATG GGA TGG AGC          579                                                                       Gly Leu Ala Asp Glu Asn Thr Asp Ile Lys As - #n Pro Met Gly Trp Ser               170              - #   175              - #   180                          - - ATT GAG GTT GGT GAT GGT ACA AAA GCC AAA AC - #T CTG ACT CTT CAG GAT          627                                                                       Ile Glu Val Gly Asp Gly Thr Lys Ala Lys Th - #r Leu Thr Leu Gln Asp           185                 1 - #90                 1 - #95                 2 -      #00                                                                              - - GTC TTG AGA CAA GGA TAC AAT ATC CTG TTT GA - #T AAC CAC AAG ATC        ACC      675                                                                    Val Leu Arg Gln Gly Tyr Asn Ile Leu Phe As - #p Asn His Lys Ile Thr                          205  - #               210  - #               215              - - TTC CAG GTG TCA TTC AAT GCC ACT GGA GTG AC - #T CAC TAC ATG CAA GGT          723                                                                       Phe Gln Val Ser Phe Asn Ala Thr Gly Val Th - #r His Tyr Met Gln Gly                       220      - #           225      - #           230                  - - AAC AGT CAC CTC TAC ATG GTG CCT CTG AAG TT - #G ATA CAT GAA TCT CTT          771                                                                       Asn Ser His Leu Tyr Met Val Pro Leu Lys Le - #u Ile His Glu Ser Leu                   235          - #       240          - #       245                      - - GGG CAG AAG ATC ATC TTA ACA ACA CGA GTG CT - #T TGT ATG TCA GAT GCT          819                                                                       Gly Gln Lys Ile Ile Leu Thr Thr Arg Val Le - #u Cys Met Ser Asp Ala               250              - #   255              - #   260                          - - GTG ACC TGT AAT GCC ACA CAT GTG ACT CTG AC - #C ATA CCA GAG TTT CCT          867                                                                       Val Thr Cys Asn Ala Thr His Val Thr Leu Th - #r Ile Pro Glu Phe Pro           265                 2 - #70                 2 - #75                 2 -      #80                                                                              - - GGG AAG TTA AAA TCT GTG AGC TCT GAA AAT AG - #G AAC TTT GCT GTA        AGC      915                                                                    Gly Lys Leu Lys Ser Val Ser Ser Glu Asn Ar - #g Asn Phe Ala Val Ser                          285  - #               290  - #               295              - - CAG CTG CAC AAC AAT GGG ATT GAT AAA GAA GA - #A TCA AGT GGC TTG ACA          963                                                                       Gln Leu His Asn Asn Gly Ile Asp Lys Glu Gl - #u Ser Ser Gly Leu Thr                       300      - #           305      - #           310                  - - TTG CAC TTC AGC AAA ACT CTT CTC AAA ATG GA - #A TTC TCT GAA AAA TGC         1011                                                                       Leu His Phe Ser Lys Thr Leu Leu Lys Met Gl - #u Phe Ser Glu Lys Cys                   315          - #       320          - #       325                      - - CTA CCC TAT CAG TTC TAC TTA GCT TCA CTC AA - #G CTG ACC TTT GCC TTT         1059                                                                       Leu Pro Tyr Gln Phe Tyr Leu Ala Ser Leu Ly - #s Leu Thr Phe Ala Phe               330              - #   335              - #   340                          - - AAT CAA GAG ACT ATA TCC ACG GTG CTT TAT CC - #T GAG TGT GTC TGT GAG         1107                                                                       Asn Gln Glu Thr Ile Ser Thr Val Leu Tyr Pr - #o Glu Cys Val Cys Glu           345                 3 - #50                 3 - #55                 3 -      #60                                                                              - - TCA CCA GTT TCT ATA GTT ACA GGT GAC CTG TG - #T ACT CAG GAT GGG        TTT     1155                                                                    Ser Pro Val Ser Ile Val Thr Gly Asp Leu Cy - #s Thr Gln Asp Gly Phe                          365  - #               370  - #               375              - - ATG GAC ATA AAG GTC TAC AGT CAC CAG ACA AA - #A CCA GCT CTC AAC TTA         1203                                                                       Met Asp Ile Lys Val Tyr Ser His Gln Thr Ly - #s Pro Ala Leu Asn Leu                       380      - #           385      - #           390                  - - GAA ACC CTA AGG GTG GGA GAC TCA TCC TGC CA - #A CCT ACC TTC CAG GCT         1251                                                                       Glu Thr Leu Arg Val Gly Asp Ser Ser Cys Gl - #n Pro Thr Phe Gln Ala                   395          - #       400          - #       405                      - - GCA TCT CAA GGG CTG ATA CTG TTT CAC ATA CC - #C CTG AAT GGA TGC GGG         1299                                                                       Ala Ser Gln Gly Leu Ile Leu Phe His Ile Pr - #o Leu Asn Gly Cys Gly               410              - #   415              - #   420                          - - ACA AGA CAT AAG TTC AAG GAA GGC AAA GTC AT - #C TAT GAA AAT GAA ATA         1347                                                                       Thr Arg His Lys Phe Lys Glu Gly Lys Val Il - #e Tyr Glu Asn Glu Ile           425                 4 - #30                 4 - #35                 4 -      #40                                                                              - - CAT GCT GTC TGG GCG GAT CTT CCT CCA AGC AC - #A ATT TCT AGA GAT        AGT     1395                                                                    His Ala Val Trp Ala Asp Leu Pro Pro Ser Th - #r Ile Ser Arg Asp Ser                          445  - #               450  - #               455              - - GAA TTC AGA ATG ACA GTG CAG TGC CAT TAC AG - #C AAA GGT GAC CTG CTA         1443                                                                       Glu Phe Arg Met Thr Val Gln Cys His Tyr Se - #r Lys Gly Asp Leu Leu                       460      - #           465      - #           470                  - - ATA AAT ACC AGA GTC CAA AGT CTT CCT CCT CT - #A GAG GCC TCA GTG AGG         1491                                                                       Ile Asn Thr Arg Val Gln Ser Leu Pro Pro Le - #u Glu Ala Ser Val Arg                   475          - #       480          - #       485                      - - CCA GGT CCA CTT GCC TTA ATC CTG CAA ACC TA - #C CCA GAT AAA TCC TAC         1539                                                                       Pro Gly Pro Leu Ala Leu Ile Leu Gln Thr Ty - #r Pro Asp Lys Ser Tyr               490              - #   495              - #   500                          - - CTC CAA CCT TAC GGG GAG AAG GAG TAC CCT GT - #G GTG AGA TAC CTC CGC         1587                                                                       Leu Gln Pro Tyr Gly Glu Lys Glu Tyr Pro Va - #l Val Arg Tyr Leu Arg           505                 5 - #10                 5 - #15                 5 -      #20                                                                              - - CAA CCA ATT TAT CTG GAA GTG AGA GTC CTA AA - #T AGG TCT GAC CCC        AAC     1635                                                                    Gln Pro Ile Tyr Leu Glu Val Arg Val Leu As - #n Arg Ser Asp Pro Asn                          525  - #               530  - #               535              - - ATC AAG CTG GTC TTA GAT GAC TGC TGG GCA AC - #A CCC ACG ATG GAC CCA         1683                                                                       Ile Lys Leu Val Leu Asp Asp Cys Trp Ala Th - #r Pro Thr Met Asp Pro                       540      - #           545      - #           550                  - - GCC TCC GTC CCC CAG TGG AAT ATT ATC ATG GA - #T GGC TGT GAA TAC AAC         1731                                                                       Ala Ser Val Pro Gln Trp Asn Ile Ile Met As - #p Gly Cys Glu Tyr Asn                   555          - #       560          - #       565                      - - CTG GAC AAC CAC AGA ACC ACC TTC CAT CCA GT - #T GGC TCC TCT GTG ACC         1779                                                                       Leu Asp Asn His Arg Thr Thr Phe His Pro Va - #l Gly Ser Ser Val Thr               570              - #   575              - #   580                          - - TAT CCT ACT CAC TAT CGG AGG TTT GAT GTG AA - #G ACC TTT GCC TTT GTA         1827                                                                       Tyr Pro Thr His Tyr Arg Arg Phe Asp Val Ly - #s Thr Phe Ala Phe Val           585                 5 - #90                 5 - #95                 6 -      #00                                                                              - - TCA GAG GCC CAA GTG CTT TCT AGT CTG GTC TA - #C TTC CAC TGC AGT        GTC     1875                                                                    Ser Glu Ala Gln Val Leu Ser Ser Leu Val Ty - #r Phe His Cys Ser Val                          605  - #               610  - #               615              - - TTA ATC TGC AGT CGA CTG TCT GCT GAC TCC CC - #T CTG TGT TCC GTG ACT         1923                                                                       Leu Ile Cys Ser Arg Leu Ser Ala Asp Ser Pr - #o Leu Cys Ser Val Thr                       620      - #           625      - #           630                  - - TGC CCT GTG TCA TTC AGA CAC AGG AGA GCC AC - #A GGC ACC ACT GAA GAA         1971                                                                       Cys Pro Val Ser Phe Arg His Arg Arg Ala Th - #r Gly Thr Thr Glu Glu                   635          - #       640          - #       645                      - - GAG AAA ATG ATA GTG AGT CTT CCA GGA CCC AT - #C CTC CTG CTG TCA GAT         2019                                                                       Glu Lys Met Ile Val Ser Leu Pro Gly Pro Il - #e Leu Leu Leu Ser Asp               650              - #   655              - #   660                          - - AGC TCT TCA CTC AGA GAT GTG GTG GAC TCA AA - #A GGG TAT GGG GCT GCC         2067                                                                       Ser Ser Ser Leu Arg Asp Val Val Asp Ser Ly - #s Gly Tyr Gly Ala Ala           665                 6 - #70                 6 - #75                 6 -      #80                                                                              - - GGA TAT GTT GCT TTT AAG ACT GTG GTA GCT GT - #G GCT GCC TTA GCA        GGC     2115                                                                    Gly Tyr Val Ala Phe Lys Thr Val Val Ala Va - #l Ala Ala Leu Ala Gly                          685  - #               690  - #               695              - - CTC GTG GCA ACG CTA GGC TTC ATC ACC TAC CT - #G CGC AAG AAC AGA ACC         2163                                                                       Leu Val Ala Thr Leu Gly Phe Ile Thr Tyr Le - #u Arg Lys Asn Arg Thr                       700      - #           705      - #           710                  - - ATG ATA AAT CAC TAAGGATTTT CAAATAAAAT GGTTGAAGTA AA - #AAAAAAAA             2215                                                                       Met Ile Asn His                                                                       715                                                                    - - AAAAAAAGCG GCCGCGAATT C           - #                  - #                    2236                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:14:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 716 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                              - - Met Ala Ser Arg Gln Lys Gly Asp Ser Gly Se - #r Pro Ser Ser Trp Phe        1               5 - #                 10 - #                 15              - - Asn Ala Asp Trp Ser Thr Tyr Arg Ser Leu Ph - #e Leu Leu Phe Ile Leu                   20     - #             25     - #             30                  - - Val Thr Ser Val Asn Ser Ile Gly Val Leu Gl - #n Leu Val Asn Pro Val               35         - #         40         - #         45                      - - Phe Pro Gly Thr Val Thr Cys Tyr Glu Thr Ar - #g Met Ala Val Glu Phe           50             - #     55             - #     60                          - - Pro Ser Asp Phe Gly Thr Lys Lys Trp His Th - #r Ser Val Val Asp Pro       65                 - # 70                 - # 75                 - # 80       - - Phe Ser Phe Glu Leu Leu Asn Cys Thr Tyr Il - #e Leu Asp Pro Glu Asn                       85 - #                 90 - #                 95              - - Leu Thr Leu Lys Ala Pro Tyr Glu Thr Cys Th - #r Arg Arg Thr Leu Gly                  100      - #           105      - #           110                  - - Gln His Arg Met Ile Ile Arg Leu Lys Asp Hi - #s Asn Ala Ala Ser Arg              115          - #       120          - #       125                      - - His Asn Ser Leu Met Tyr Gln Ile Asn Cys Pr - #o Val Met Gln Ala Glu          130              - #   135              - #   140                          - - Glu Thr His Glu His Ala Gly Ser Thr Ile Cy - #s Thr Lys Asp Ser Met      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ser Phe Thr Phe Asn Val Ile Pro Gly Leu Al - #a Asp Glu Asn Thr        Asp                                                                                             165  - #               170  - #               175             - - Ile Lys Asn Pro Met Gly Trp Ser Ile Glu Va - #l Gly Asp Gly Thr Lys                  180      - #           185      - #           190                  - - Ala Lys Thr Leu Thr Leu Gln Asp Val Leu Ar - #g Gln Gly Tyr Asn Ile              195          - #       200          - #       205                      - - Leu Phe Asp Asn His Lys Ile Thr Phe Gln Va - #l Ser Phe Asn Ala Thr          210              - #   215              - #   220                          - - Gly Val Thr His Tyr Met Gln Gly Asn Ser Hi - #s Leu Tyr Met Val Pro      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Leu Lys Leu Ile His Glu Ser Leu Gly Gln Ly - #s Ile Ile Leu Thr        Thr                                                                                             245  - #               250  - #               255             - - Arg Val Leu Cys Met Ser Asp Ala Val Thr Cy - #s Asn Ala Thr His Val                  260      - #           265      - #           270                  - - Thr Leu Thr Ile Pro Glu Phe Pro Gly Lys Le - #u Lys Ser Val Ser Ser              275          - #       280          - #       285                      - - Glu Asn Arg Asn Phe Ala Val Ser Gln Leu Hi - #s Asn Asn Gly Ile Asp          290              - #   295              - #   300                          - - Lys Glu Glu Ser Ser Gly Leu Thr Leu His Ph - #e Ser Lys Thr Leu Leu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Lys Met Glu Phe Ser Glu Lys Cys Leu Pro Ty - #r Gln Phe Tyr Leu        Ala                                                                                             325  - #               330  - #               335             - - Ser Leu Lys Leu Thr Phe Ala Phe Asn Gln Gl - #u Thr Ile Ser Thr Val                  340      - #           345      - #           350                  - - Leu Tyr Pro Glu Cys Val Cys Glu Ser Pro Va - #l Ser Ile Val Thr Gly              355          - #       360          - #       365                      - - Asp Leu Cys Thr Gln Asp Gly Phe Met Asp Il - #e Lys Val Tyr Ser His          370              - #   375              - #   380                          - - Gln Thr Lys Pro Ala Leu Asn Leu Glu Thr Le - #u Arg Val Gly Asp Ser      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Cys Gln Pro Thr Phe Gln Ala Ala Ser Gl - #n Gly Leu Ile Leu        Phe                                                                                             405  - #               410  - #               415             - - His Ile Pro Leu Asn Gly Cys Gly Thr Arg Hi - #s Lys Phe Lys Glu Gly                  420      - #           425      - #           430                  - - Lys Val Ile Tyr Glu Asn Glu Ile His Ala Va - #l Trp Ala Asp Leu Pro              435          - #       440          - #       445                      - - Pro Ser Thr Ile Ser Arg Asp Ser Glu Phe Ar - #g Met Thr Val Gln Cys          450              - #   455              - #   460                          - - His Tyr Ser Lys Gly Asp Leu Leu Ile Asn Th - #r Arg Val Gln Ser Leu      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Pro Pro Leu Glu Ala Ser Val Arg Pro Gly Pr - #o Leu Ala Leu Ile        Leu                                                                                             485  - #               490  - #               495             - - Gln Thr Tyr Pro Asp Lys Ser Tyr Leu Gln Pr - #o Tyr Gly Glu Lys Glu                  500      - #           505      - #           510                  - - Tyr Pro Val Val Arg Tyr Leu Arg Gln Pro Il - #e Tyr Leu Glu Val Arg              515          - #       520          - #       525                      - - Val Leu Asn Arg Ser Asp Pro Asn Ile Lys Le - #u Val Leu Asp Asp Cys          530              - #   535              - #   540                          - - Trp Ala Thr Pro Thr Met Asp Pro Ala Ser Va - #l Pro Gln Trp Asn Ile      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Ile Met Asp Gly Cys Glu Tyr Asn Leu Asp As - #n His Arg Thr Thr        Phe                                                                                             565  - #               570  - #               575             - - His Pro Val Gly Ser Ser Val Thr Tyr Pro Th - #r His Tyr Arg Arg Phe                  580      - #           585      - #           590                  - - Asp Val Lys Thr Phe Ala Phe Val Ser Glu Al - #a Gln Val Leu Ser Ser              595          - #       600          - #       605                      - - Leu Val Tyr Phe His Cys Ser Val Leu Ile Cy - #s Ser Arg Leu Ser Ala          610              - #   615              - #   620                          - - Asp Ser Pro Leu Cys Ser Val Thr Cys Pro Va - #l Ser Phe Arg His Arg      625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - Arg Ala Thr Gly Thr Thr Glu Glu Glu Lys Me - #t Ile Val Ser Leu        Pro                                                                                             645  - #               650  - #               655             - - Gly Pro Ile Leu Leu Leu Ser Asp Ser Ser Se - #r Leu Arg Asp Val Val                  660      - #           665      - #           670                  - - Asp Ser Lys Gly Tyr Gly Ala Ala Gly Tyr Va - #l Ala Phe Lys Thr Val              675          - #       680          - #       685                      - - Val Ala Val Ala Ala Leu Ala Gly Leu Val Al - #a Thr Leu Gly Phe Ile          690              - #   695              - #   700                          - - Thr Tyr Leu Arg Lys Asn Arg Thr Met Ile As - #n His                      705                 7 - #10                 7 - #15                            - -  - - (2) INFORMATION FOR SEQ ID NO:15:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1840 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Felis dom - #esticus                                            (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 57..1766                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                              - - GAATTCCGCG GCCGCAAGTA CAGGTCTTGC AGCCAGTGGG GGCTCCCGAT GG - #CATC             56                                                                        - - ATG TGG CTG CTG CAG CCC CTC TTG CTC TGT GT - #T CCC TTG TCT CTC GCT          104                                                                       Met Trp Leu Leu Gln Pro Leu Leu Leu Cys Va - #l Pro Leu Ser Leu Ala             1               5 - #                 10 - #                 15              - - GTG CAT GGC CAG CAG AAG CCC CAG GTA CCA GA - #T TAT CCC GGT GAA CTC          152                                                                       Val His Gly Gln Gln Lys Pro Gln Val Pro As - #p Tyr Pro Gly Glu Leu                        20     - #             25     - #             30                  - - CAT TGT GGG CTC CAG AGC CTT CAG TTT GCC AT - #A AAC CCG AGC CCC GGG          200                                                                       His Cys Gly Leu Gln Ser Leu Gln Phe Ala Il - #e Asn Pro Ser Pro Gly                    35         - #         40         - #         45                      - - AAA GCG ACT CCT GCA CTC ATA GTC TGG GAC AA - #T CGC GGG CTG CCA CAC          248                                                                       Lys Ala Thr Pro Ala Leu Ile Val Trp Asp As - #n Arg Gly Leu Pro His                50             - #     55             - #     60                          - - AAG CTG CAG AAC AAC TCT GGC TGC GGT ACC TG - #G GTA AGG GAG AGC CCG          296                                                                       Lys Leu Gln Asn Asn Ser Gly Cys Gly Thr Tr - #p Val Arg Glu Ser Pro            65                 - # 70                 - # 75                 - # 80       - - GGG GGC TCC GTG CTG TTA GAC GCC TCT TAC AG - #C AGC TGC TAT GTC AAC          344                                                                       Gly Gly Ser Val Leu Leu Asp Ala Ser Tyr Se - #r Ser Cys Tyr Val Asn                            85 - #                 90 - #                 95              - - GAG TGG GTG AGC ACG ACC CAA TCC CCA GGA AC - #G TCG AGG CCC CCC ACC          392                                                                       Glu Trp Val Ser Thr Thr Gln Ser Pro Gly Th - #r Ser Arg Pro Pro Thr                       100      - #           105      - #           110                  - - CCA GCA TCC AGG GTG ACT CCC CAG GAC TCC CA - #C TAC GTC ATG ATA GTC          440                                                                       Pro Ala Ser Arg Val Thr Pro Gln Asp Ser Hi - #s Tyr Val Met Ile Val                   115          - #       120          - #       125                      - - GGA GTT GAA GGC ACA GAT GCG GCT GGG CGC AG - #G GTT ACC AAC ACC AAG          488                                                                       Gly Val Glu Gly Thr Asp Ala Ala Gly Arg Ar - #g Val Thr Asn Thr Lys               130              - #   135              - #   140                          - - GTG CTC AGG TGT CCT AGG AAT CCC CCA GAC CA - #A GCT TTG GTG TCG AGC          536                                                                       Val Leu Arg Cys Pro Arg Asn Pro Pro Asp Gl - #n Ala Leu Val Ser Ser           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - TTA AGT CCC TCT CCT CTT CAA AAC GTA GCA CT - #A GAA GCT CCA AAC        GCT      584                                                                    Leu Ser Pro Ser Pro Leu Gln Asn Val Ala Le - #u Glu Ala Pro Asn Ala                          165  - #               170  - #               175              - - GAC TTG TGT GAC TCT GTC CCA AAG TGG GAC AG - #G CTT CCG TGT GCT TCT          632                                                                       Asp Leu Cys Asp Ser Val Pro Lys Trp Asp Ar - #g Leu Pro Cys Ala Ser                       180      - #           185      - #           190                  - - TCA CCC ATC ACT CAG GGA GAC TGC AAT AAG CT - #T GGT TGC TGC TAC AAA          680                                                                       Ser Pro Ile Thr Gln Gly Asp Cys Asn Lys Le - #u Gly Cys Cys Tyr Lys                   195          - #       200          - #       205                      - - TCA GAG GCA AAT TCC TGT TAC TAT GGA AAC AC - #A GTG ACC TCA CGC TGT          728                                                                       Ser Glu Ala Asn Ser Cys Tyr Tyr Gly Asn Th - #r Val Thr Ser Arg Cys               210              - #   215              - #   220                          - - ACC CAA GAC GGC CAC TTC TCC ATC GCC GTG TC - #T CGG AAC GTG ACC TCA          776                                                                       Thr Gln Asp Gly His Phe Ser Ile Ala Val Se - #r Arg Asn Val Thr Ser           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - CCC CCA CTG CTC TTA AAT TCT CTG CGC TTG GC - #C TTC GGG AAG GAC        CGC      824                                                                    Pro Pro Leu Leu Leu Asn Ser Leu Arg Leu Al - #a Phe Gly Lys Asp Arg                          245  - #               250  - #               255              - - GAA TGT AAC CCT GTG AAA GCA ACA CGT GCC TT - #T GCC CTG TTC TTT TTT          872                                                                       Glu Cys Asn Pro Val Lys Ala Thr Arg Ala Ph - #e Ala Leu Phe Phe Phe                       260      - #           265      - #           270                  - - CCA TTT AAT TCC TGT GGC ACC ACG AGA TGG GT - #C ACT GGA GAC CAG GCA          920                                                                       Pro Phe Asn Ser Cys Gly Thr Thr Arg Trp Va - #l Thr Gly Asp Gln Ala                   275          - #       280          - #       285                      - - GTA TAT GAA AAT GAG CTG GTG GCA GCT AGA GA - #T GTG AGA ACT TGG AGC          968                                                                       Val Tyr Glu Asn Glu Leu Val Ala Ala Arg As - #p Val Arg Thr Trp Ser               290              - #   295              - #   300                          - - CAT GGT TCT ATT ACC CGT GAC AGT ATC TTC AG - #G CTT CGA GTT AGC TGC         1016                                                                       His Gly Ser Ile Thr Arg Asp Ser Ile Phe Ar - #g Leu Arg Val Ser Cys           305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - AGC TAC TCT GTA AGG AGT AAT GCC TTC CCG CT - #T AGC GTT CAG GTG        TTT     1064                                                                    Ser Tyr Ser Val Arg Ser Asn Ala Phe Pro Le - #u Ser Val Gln Val Phe                          325  - #               330  - #               335              - - ACC ATC CCA CCA CCC CAT CTG AAA ACC CAG CA - #T GGA CCC CTC ACT CTG         1112                                                                       Thr Ile Pro Pro Pro His Leu Lys Thr Gln Hi - #s Gly Pro Leu Thr Leu                       340      - #           345      - #           350                  - - GAA CTC AAG ATT GCC AAA GAT AAG CAC TAT GG - #C TCC TAC TAC ACT ATT         1160                                                                       Glu Leu Lys Ile Ala Lys Asp Lys His Tyr Gl - #y Ser Tyr Tyr Thr Ile                   355          - #       360          - #       365                      - - GGT GAC TAC CCA GTG GTA AAG TTG CTT CGG GA - #T CCC ATT TAT GTG GAG         1208                                                                       Gly Asp Tyr Pro Val Val Lys Leu Leu Arg As - #p Pro Ile Tyr Val Glu               370              - #   375              - #   380                          - - GTC TCT ATC CGC CAC AGA ACG GAC CCC TCC CT - #G GGG CTG CTC CTC CAT         1256                                                                       Val Ser Ile Arg His Arg Thr Asp Pro Ser Le - #u Gly Leu Leu Leu His           385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - AAC TGT TGG GCC ACA CCC GGC AAG AAC TCC CA - #G AGT CTG TCC CAG        TGG     1304                                                                    Asn Cys Trp Ala Thr Pro Gly Lys Asn Ser Gl - #n Ser Leu Ser Gln Trp                          405  - #               410  - #               415              - - CCC ATT CTG GTG AAA GGA TGC CCC TAC GTT GG - #A GAC AAC TAT CAA ACC         1352                                                                       Pro Ile Leu Val Lys Gly Cys Pro Tyr Val Gl - #y Asp Asn Tyr Gln Thr                       420      - #           425      - #           430                  - - CAG CTG ATC CCT GTC CAG AAG GCT CTG GAT AC - #A CCA TTT CCA TCT TAC         1400                                                                       Gln Leu Ile Pro Val Gln Lys Ala Leu Asp Th - #r Pro Phe Pro Ser Tyr                   435          - #       440          - #       445                      - - TAC AAG CGC TTC AGT ATT TTC ACC TTC AGC TT - #T GTG GAC ACC ATG GCA         1448                                                                       Tyr Lys Arg Phe Ser Ile Phe Thr Phe Ser Ph - #e Val Asp Thr Met Ala               450              - #   455              - #   460                          - - AAG TGG GCA CTC AGG GGA CCG GTG TAT CTG CA - #C TGT AAT GTA TCC ATC         1496                                                                       Lys Trp Ala Leu Arg Gly Pro Val Tyr Leu Hi - #s Cys Asn Val Ser Ile           465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - TGC CAG CCT GCT GGG ACC TCC TCC TGT AGG AT - #A ACC TGT CCT GTT        GCC     1544                                                                    Cys Gln Pro Ala Gly Thr Ser Ser Cys Arg Il - #e Thr Cys Pro Val Ala                          485  - #               490  - #               495              - - AGG CGA AGA AGA CAC TCT GAC CTC CAT CAT CA - #C AGC AGT ACT GCG AGC         1592                                                                       Arg Arg Arg Arg His Ser Asp Leu His His Hi - #s Ser Ser Thr Ala Ser                       500      - #           505      - #           510                  - - ATC TCT AGC AAG GGT CCC ATG ATT CTA CTC CA - #A GCC ACT ATG GAC TCT         1640                                                                       Ile Ser Ser Lys Gly Pro Met Ile Leu Leu Gl - #n Ala Thr Met Asp Ser                   515          - #       520          - #       525                      - - GCA GAG AAG CTC CAC AAA AAC TCA AGT TCT CC - #T ATA GAC TCC CAA GCT         1688                                                                       Ala Glu Lys Leu His Lys Asn Ser Ser Ser Pr - #o Ile Asp Ser Gln Ala               530              - #   535              - #   540                          - - CTG TGG ATG GCA GGC CTT TCC GGG ACC CTA AT - #C TTT GGA TTC TTG TTA         1736                                                                       Leu Trp Met Ala Gly Leu Ser Gly Thr Leu Il - #e Phe Gly Phe Leu Leu           545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - GTG TCC TAC TTG GCT ATC AGG AAA CGG AGG TG - #AATTATTC CAGTTGTGTT           1786                                                                      Val Ser Tyr Leu Ala Ile Arg Lys Arg Arg                                                       565  - #               570                                     - - AATAAAACCA GATTGCATTA CCAAAAAAAA AAAAAAAAAA GCGGCCGCGA AT - #TC             1840                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:16:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 570 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                              - - Met Trp Leu Leu Gln Pro Leu Leu Leu Cys Va - #l Pro Leu Ser Leu Ala        1               5 - #                 10 - #                 15              - - Val His Gly Gln Gln Lys Pro Gln Val Pro As - #p Tyr Pro Gly Glu Leu                   20     - #             25     - #             30                  - - His Cys Gly Leu Gln Ser Leu Gln Phe Ala Il - #e Asn Pro Ser Pro Gly               35         - #         40         - #         45                      - - Lys Ala Thr Pro Ala Leu Ile Val Trp Asp As - #n Arg Gly Leu Pro His           50             - #     55             - #     60                          - - Lys Leu Gln Asn Asn Ser Gly Cys Gly Thr Tr - #p Val Arg Glu Ser Pro       65                 - # 70                 - # 75                 - # 80       - - Gly Gly Ser Val Leu Leu Asp Ala Ser Tyr Se - #r Ser Cys Tyr Val Asn                       85 - #                 90 - #                 95              - - Glu Trp Val Ser Thr Thr Gln Ser Pro Gly Th - #r Ser Arg Pro Pro Thr                  100      - #           105      - #           110                  - - Pro Ala Ser Arg Val Thr Pro Gln Asp Ser Hi - #s Tyr Val Met Ile Val              115          - #       120          - #       125                      - - Gly Val Glu Gly Thr Asp Ala Ala Gly Arg Ar - #g Val Thr Asn Thr Lys          130              - #   135              - #   140                          - - Val Leu Arg Cys Pro Arg Asn Pro Pro Asp Gl - #n Ala Leu Val Ser Ser      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Leu Ser Pro Ser Pro Leu Gln Asn Val Ala Le - #u Glu Ala Pro Asn        Ala                                                                                             165  - #               170  - #               175             - - Asp Leu Cys Asp Ser Val Pro Lys Trp Asp Ar - #g Leu Pro Cys Ala Ser                  180      - #           185      - #           190                  - - Ser Pro Ile Thr Gln Gly Asp Cys Asn Lys Le - #u Gly Cys Cys Tyr Lys              195          - #       200          - #       205                      - - Ser Glu Ala Asn Ser Cys Tyr Tyr Gly Asn Th - #r Val Thr Ser Arg Cys          210              - #   215              - #   220                          - - Thr Gln Asp Gly His Phe Ser Ile Ala Val Se - #r Arg Asn Val Thr Ser      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Pro Pro Leu Leu Leu Asn Ser Leu Arg Leu Al - #a Phe Gly Lys Asp        Arg                                                                                             245  - #               250  - #               255             - - Glu Cys Asn Pro Val Lys Ala Thr Arg Ala Ph - #e Ala Leu Phe Phe Phe                  260      - #           265      - #           270                  - - Pro Phe Asn Ser Cys Gly Thr Thr Arg Trp Va - #l Thr Gly Asp Gln Ala              275          - #       280          - #       285                      - - Val Tyr Glu Asn Glu Leu Val Ala Ala Arg As - #p Val Arg Thr Trp Ser          290              - #   295              - #   300                          - - His Gly Ser Ile Thr Arg Asp Ser Ile Phe Ar - #g Leu Arg Val Ser Cys      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Ser Tyr Ser Val Arg Ser Asn Ala Phe Pro Le - #u Ser Val Gln Val        Phe                                                                                             325  - #               330  - #               335             - - Thr Ile Pro Pro Pro His Leu Lys Thr Gln Hi - #s Gly Pro Leu Thr Leu                  340      - #           345      - #           350                  - - Glu Leu Lys Ile Ala Lys Asp Lys His Tyr Gl - #y Ser Tyr Tyr Thr Ile              355          - #       360          - #       365                      - - Gly Asp Tyr Pro Val Val Lys Leu Leu Arg As - #p Pro Ile Tyr Val Glu          370              - #   375              - #   380                          - - Val Ser Ile Arg His Arg Thr Asp Pro Ser Le - #u Gly Leu Leu Leu His      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Asn Cys Trp Ala Thr Pro Gly Lys Asn Ser Gl - #n Ser Leu Ser Gln        Trp                                                                                             405  - #               410  - #               415             - - Pro Ile Leu Val Lys Gly Cys Pro Tyr Val Gl - #y Asp Asn Tyr Gln Thr                  420      - #           425      - #           430                  - - Gln Leu Ile Pro Val Gln Lys Ala Leu Asp Th - #r Pro Phe Pro Ser Tyr              435          - #       440          - #       445                      - - Tyr Lys Arg Phe Ser Ile Phe Thr Phe Ser Ph - #e Val Asp Thr Met Ala          450              - #   455              - #   460                          - - Lys Trp Ala Leu Arg Gly Pro Val Tyr Leu Hi - #s Cys Asn Val Ser Ile      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Cys Gln Pro Ala Gly Thr Ser Ser Cys Arg Il - #e Thr Cys Pro Val        Ala                                                                                             485  - #               490  - #               495             - - Arg Arg Arg Arg His Ser Asp Leu His His Hi - #s Ser Ser Thr Ala Ser                  500      - #           505      - #           510                  - - Ile Ser Ser Lys Gly Pro Met Ile Leu Leu Gl - #n Ala Thr Met Asp Ser              515          - #       520          - #       525                      - - Ala Glu Lys Leu His Lys Asn Ser Ser Ser Pr - #o Ile Asp Ser Gln Ala          530              - #   535              - #   540                          - - Leu Trp Met Ala Gly Leu Ser Gly Thr Leu Il - #e Phe Gly Phe Leu Leu      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Val Ser Tyr Leu Ala Ile Arg Lys Arg Arg                                                  565  - #               570                                     - -  - - (2) INFORMATION FOR SEQ ID NO:17:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1319 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Felis dom - #esticus                                            (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 26..1297                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                              - - GAATTCGCGG CCGCGCGTAG GCCGC ATG GGG CTG AGC TAC - #GGG CTT TTC ATC           52                                                                                         - #          Met Gly Leu Ser Tyr - # Gly Leu Phe Ile                          - #            1      - #         5                          - - TGT TTT CTG CTT TGG GCA GGC ACG GGG CTG TG - #C TAT CCC CCA ACC ACC          100                                                                       Cys Phe Leu Leu Trp Ala Gly Thr Gly Leu Cy - #s Tyr Pro Pro Thr Thr            10                 - # 15                 - # 20                 - # 25       - - ACC GAG GAT AAG ACC CAC CCC TCG TTG CCA TC - #A AGC CCC TCT GTG GTG          148                                                                       Thr Glu Asp Lys Thr His Pro Ser Leu Pro Se - #r Ser Pro Ser Val Val                            30 - #                 35 - #                 40              - - GTA GAG TGT CGG CAT GCC TGG CTG GTG GTC AA - #C GTC AGC AAA AAC CTT          196                                                                       Val Glu Cys Arg His Ala Trp Leu Val Val As - #n Val Ser Lys Asn Leu                        45     - #             50     - #             55                  - - TTT GGT ACT GGG AGG CTT GTG AGG CCT GCA GA - #C CTC ACC CTG GGT CCG          244                                                                       Phe Gly Thr Gly Arg Leu Val Arg Pro Ala As - #p Leu Thr Leu Gly Pro                    60         - #         65         - #         70                      - - GAG AAC TGT GAG CCC CTG ATC TCT GGG GAC TC - #A GAT GAT ACG GTC AGG          292                                                                       Glu Asn Cys Glu Pro Leu Ile Ser Gly Asp Se - #r Asp Asp Thr Val Arg                75             - #     80             - #     85                          - - TTT GAA GTC GAG CTC CAC AAG TGT GGC AAC AG - #C GTG CAG GTG ACC GAA          340                                                                       Phe Glu Val Glu Leu His Lys Cys Gly Asn Se - #r Val Gln Val Thr Glu            90                 - # 95                 - #100                 - #105       - - GAT GCC CTG GTG TAT AGC ACC TTC CTG CTC CA - #C AAC CCC CGC CCC ATG          388                                                                       Asp Ala Leu Val Tyr Ser Thr Phe Leu Leu Hi - #s Asn Pro Arg Pro Met                           110  - #               115  - #               120              - - GGA AAC CTG TCC ATC CTG AGG ACC AAC CGC GC - #G GAA GTT CCC ATT GAG          436                                                                       Gly Asn Leu Ser Ile Leu Arg Thr Asn Arg Al - #a Glu Val Pro Ile Glu                       125      - #           130      - #           135                  - - TGC CGT TAC CCC AGG CAT AGC AAC GTG AGC AG - #C GAG GCC ATC CTG CCC          484                                                                       Cys Arg Tyr Pro Arg His Ser Asn Val Ser Se - #r Glu Ala Ile Leu Pro                   140          - #       145          - #       150                      - - ACC TGG GTG CCC TTC AGG ACC ACA ATG CTC TC - #A GAG GAG AAG CTG GCT          532                                                                       Thr Trp Val Pro Phe Arg Thr Thr Met Leu Se - #r Glu Glu Lys Leu Ala               155              - #   160              - #   165                          - - TTC TCT CTG CGC CTG ATG GAG GAG GAC TGG GG - #C TCC GAG AAG CAG TCC          580                                                                       Phe Ser Leu Arg Leu Met Glu Glu Asp Trp Gl - #y Ser Glu Lys Gln Ser           170                 1 - #75                 1 - #80                 1 -      #85                                                                              - - CCC ACT TTC CAG TTG GGA GAC CTA GCC CAC CT - #C CAG GCC GAA GTC        CAC      628                                                                    Pro Thr Phe Gln Leu Gly Asp Leu Ala His Le - #u Gln Ala Glu Val His                          190  - #               195  - #               200              - - ACC GGC CGC CAC ATA CCA CTG CGA CTG TTT GT - #G GAC TAC TGT GTG GCC          676                                                                       Thr Gly Arg His Ile Pro Leu Arg Leu Phe Va - #l Asp Tyr Cys Val Ala                       205      - #           210      - #           215                  - - ACG CTG ACA CCA GAC CAG AAC GCC TCC CCT CA - #T CAC ACC ATC GTG GAC          724                                                                       Thr Leu Thr Pro Asp Gln Asn Ala Ser Pro Hi - #s His Thr Ile Val Asp                   220          - #       225          - #       230                      - - TTC CAC GGC TGT CTC GTG GAT GGT CTC TCT GA - #T GCC TCT TCT GCC TTC          772                                                                       Phe His Gly Cys Leu Val Asp Gly Leu Ser As - #p Ala Ser Ser Ala Phe               235              - #   240              - #   245                          - - AAA GCC CCC AGA CCC AGG CCG GAG ACT CTC CA - #G TTT ACA GTA GAC ACG          820                                                                       Lys Ala Pro Arg Pro Arg Pro Glu Thr Leu Gl - #n Phe Thr Val Asp Thr           250                 2 - #55                 2 - #60                 2 -      #65                                                                              - - TTC CAC TTT GCT AAT GAC CCC AGA AAC ATG AT - #C TAT ATC ACC TGC        CAT      868                                                                    Phe His Phe Ala Asn Asp Pro Arg Asn Met Il - #e Tyr Ile Thr Cys His                          270  - #               275  - #               280              - - CTG AAA GTC ACT CCA GCT AGC CGA GTC CCA GA - #C CAG CTA AAC AAA GCC          916                                                                       Leu Lys Val Thr Pro Ala Ser Arg Val Pro As - #p Gln Leu Asn Lys Ala                       285      - #           290      - #           295                  - - TGT TCC TTC ATC AAG TCT TCT AAC AGG TGG TT - #C CCA GTA GAA GGC CCT          964                                                                       Cys Ser Phe Ile Lys Ser Ser Asn Arg Trp Ph - #e Pro Val Glu Gly Pro                   300          - #       305          - #       310                      - - GCT GAC ATC TGT AAC TGT TGT AAC AAA GGT AG - #C TGT GGC CTT CAG GGC         1012                                                                       Ala Asp Ile Cys Asn Cys Cys Asn Lys Gly Se - #r Cys Gly Leu Gln Gly               315              - #   320              - #   325                          - - CGT TCC TGG AGG CTG TCC CAC CTA GAC AGA CC - #G TGG CAC AAG ATG GCT         1060                                                                       Arg Ser Trp Arg Leu Ser His Leu Asp Arg Pr - #o Trp His Lys Met Ala           330                 3 - #35                 3 - #40                 3 -      #45                                                                              - - TCC CGA AAT CGC AGG CAT GTG ACC GAA GAA GC - #G GAT ATC ACC GTG        GGG     1108                                                                    Ser Arg Asn Arg Arg His Val Thr Glu Glu Al - #a Asp Ile Thr Val Gly                          350  - #               355  - #               360              - - CCT CTG ATC TTC CTG GGA AAG GCT GCC GAT CG - #T GGT GTG GAG GGG TCG         1156                                                                       Pro Leu Ile Phe Leu Gly Lys Ala Ala Asp Ar - #g Gly Val Glu Gly Ser                       365      - #           370      - #           375                  - - ACC TCG CCT CAC ACC TCT GTG ATG GTG GGC AT - #A GGC CTG GCC ACG GTG         1204                                                                       Thr Ser Pro His Thr Ser Val Met Val Gly Il - #e Gly Leu Ala Thr Val                   380          - #       385          - #       390                      - - TTG TCC CTG ACT CTG GCT ACC ATT GTC CTG GG - #T CTC GCC AGG AGG CAT         1252                                                                       Leu Ser Leu Thr Leu Ala Thr Ile Val Leu Gl - #y Leu Ala Arg Arg His               395              - #   400              - #   405                          - - CAC ACT GCT TCC CGT CCT ATG ATC TGC CCT GT - #G TCT GCT TCC CAA             1297                                                                       His Thr Ala Ser Arg Pro Met Ile Cys Pro Va - #l Ser Ala Ser Gln               410                 4 - #15                 4 - #20                            - - TAAAAGAAGC GGCCGCGAAT TC           - #                  - #                   1319                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:18:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 424 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                              - - Met Gly Leu Ser Tyr Gly Leu Phe Ile Cys Ph - #e Leu Leu Trp Ala Gly        1               5 - #                 10 - #                 15              - - Thr Gly Leu Cys Tyr Pro Pro Thr Thr Thr Gl - #u Asp Lys Thr His Pro                   20     - #             25     - #             30                  - - Ser Leu Pro Ser Ser Pro Ser Val Val Val Gl - #u Cys Arg His Ala Trp               35         - #         40         - #         45                      - - Leu Val Val Asn Val Ser Lys Asn Leu Phe Gl - #y Thr Gly Arg Leu Val           50             - #     55             - #     60                          - - Arg Pro Ala Asp Leu Thr Leu Gly Pro Glu As - #n Cys Glu Pro Leu Ile       65                 - # 70                 - # 75                 - # 80       - - Ser Gly Asp Ser Asp Asp Thr Val Arg Phe Gl - #u Val Glu Leu His Lys                       85 - #                 90 - #                 95              - - Cys Gly Asn Ser Val Gln Val Thr Glu Asp Al - #a Leu Val Tyr Ser Thr                  100      - #           105      - #           110                  - - Phe Leu Leu His Asn Pro Arg Pro Met Gly As - #n Leu Ser Ile Leu Arg              115          - #       120          - #       125                      - - Thr Asn Arg Ala Glu Val Pro Ile Glu Cys Ar - #g Tyr Pro Arg His Ser          130              - #   135              - #   140                          - - Asn Val Ser Ser Glu Ala Ile Leu Pro Thr Tr - #p Val Pro Phe Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Thr Met Leu Ser Glu Glu Lys Leu Ala Phe Se - #r Leu Arg Leu Met        Glu                                                                                             165  - #               170  - #               175             - - Glu Asp Trp Gly Ser Glu Lys Gln Ser Pro Th - #r Phe Gln Leu Gly Asp                  180      - #           185      - #           190                  - - Leu Ala His Leu Gln Ala Glu Val His Thr Gl - #y Arg His Ile Pro Leu              195          - #       200          - #       205                      - - Arg Leu Phe Val Asp Tyr Cys Val Ala Thr Le - #u Thr Pro Asp Gln Asn          210              - #   215              - #   220                          - - Ala Ser Pro His His Thr Ile Val Asp Phe Hi - #s Gly Cys Leu Val Asp      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Gly Leu Ser Asp Ala Ser Ser Ala Phe Lys Al - #a Pro Arg Pro Arg        Pro                                                                                             245  - #               250  - #               255             - - Glu Thr Leu Gln Phe Thr Val Asp Thr Phe Hi - #s Phe Ala Asn Asp Pro                  260      - #           265      - #           270                  - - Arg Asn Met Ile Tyr Ile Thr Cys His Leu Ly - #s Val Thr Pro Ala Ser              275          - #       280          - #       285                      - - Arg Val Pro Asp Gln Leu Asn Lys Ala Cys Se - #r Phe Ile Lys Ser Ser          290              - #   295              - #   300                          - - Asn Arg Trp Phe Pro Val Glu Gly Pro Ala As - #p Ile Cys Asn Cys Cys      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Asn Lys Gly Ser Cys Gly Leu Gln Gly Arg Se - #r Trp Arg Leu Ser        His                                                                                             325  - #               330  - #               335             - - Leu Asp Arg Pro Trp His Lys Met Ala Ser Ar - #g Asn Arg Arg His Val                  340      - #           345      - #           350                  - - Thr Glu Glu Ala Asp Ile Thr Val Gly Pro Le - #u Ile Phe Leu Gly Lys              355          - #       360          - #       365                      - - Ala Ala Asp Arg Gly Val Glu Gly Ser Thr Se - #r Pro His Thr Ser Val          370              - #   375              - #   380                          - - Met Val Gly Ile Gly Leu Ala Thr Val Leu Se - #r Leu Thr Leu Ala Thr      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ile Val Leu Gly Leu Ala Arg Arg His His Th - #r Ala Ser Arg Pro        Met                                                                                             405  - #               410  - #               415             - - Ile Cys Pro Val Ser Ala Ser Gln                                                      420                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:19:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 643 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bos tauru - #s                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 16..582                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                              - - GAATTCGCGG CCGCC CTA AAC AGG ACT GAC CCC AAC - #ATC AAG TTG GTC TTA          51                                                                                         Leu - #Asn Arg Thr Asp Pro Asn Ile Lys Leu Val L - #eu                         - # 1               5  - #                10                 - - GAT GAT TGC TGG GCA ACA TCC ACC ATG GAC CC - #A GCC TCT CTC CCT CAG           99                                                                       Asp Asp Cys Trp Ala Thr Ser Thr Met Asp Pr - #o Ala Ser Leu Pro Gln                    15         - #         20         - #         25                      - - TGG AAT ATT ATC GTG GAT GGC TGT GAA TAC AA - #C TTG GAC AAC CAC AGA          147                                                                       Trp Asn Ile Ile Val Asp Gly Cys Glu Tyr As - #n Leu Asp Asn His Arg                30             - #     35             - #     40                          - - ACC ACC TTC CAT CCG GTT GGC TCC TCG GTG GC - #C TAT CCT AAT CAC TAC          195                                                                       Thr Thr Phe His Pro Val Gly Ser Ser Val Al - #a Tyr Pro Asn His Tyr            45                 - # 50                 - # 55                 - # 60       - - CAG AGG TTT GCT GTG AAG ACC TTT GCC TTT GT - #G TCA GAG GAC CCG GCG          243                                                                       Gln Arg Phe Ala Val Lys Thr Phe Ala Phe Va - #l Ser Glu Asp Pro Ala                            65 - #                 70 - #                 75              - - TTC TCT CAC TTG GTC TAC TTC CAC TGC AGC GC - #C TTA ATC TGC GAT CAA          291                                                                       Phe Ser His Leu Val Tyr Phe His Cys Ser Al - #a Leu Ile Cys Asp Gln                        80     - #             85     - #             90                  - - CTT TCT TCT AAC TTC CCC CTG TGT TCT GCG TC - #T TGC CTT GTG TCA TCC          339                                                                       Leu Ser Ser Asn Phe Pro Leu Cys Ser Ala Se - #r Cys Leu Val Ser Ser                    95         - #        100         - #        105                      - - AGA AGC AGG CGA GCC ACA GGG GCC ACT GAG GA - #A GAG AAG ATG ATA GTG          387                                                                       Arg Ser Arg Arg Ala Thr Gly Ala Thr Glu Gl - #u Glu Lys Met Ile Val               110              - #   115              - #   120                          - - AGT CTC CCG GGC CCC ATC CTC CTG TTG TCA GA - #T GGC TCT TCA TTC AGA          435                                                                       Ser Leu Pro Gly Pro Ile Leu Leu Leu Ser As - #p Gly Ser Ser Phe Arg           125                 1 - #30                 1 - #35                 1 -      #40                                                                              - - GAT GCT GTG GAT TCT AAA GGG CAT GGG ACT TC - #T GGA TAT GCT GCT        TTT      483                                                                    Asp Ala Val Asp Ser Lys Gly His Gly Thr Se - #r Gly Tyr Ala Ala Phe                          145  - #               150  - #               155              - - AAA ACT ATG GTT GCT GTA GTT GCC TTA GCA GG - #T GTT GTG GCA ACT CTA          531                                                                       Lys Thr Met Val Ala Val Val Ala Leu Ala Gl - #y Val Val Ala Thr Leu                       160      - #           165      - #           170                  - - AGC CTA ATC AGC TAC CTG CGC AAG AAA AGA AT - #C ACA GTG CTA AAC CAC          579                                                                       Ser Leu Ile Ser Tyr Leu Arg Lys Lys Arg Il - #e Thr Val Leu Asn His                   175          - #       180          - #       185                      - - TAATTGGATT TTCAATAAAA TGTGGAAGTA AAAAAAAAAA AAAAAAAAAA GC -             #GGCCGCGA    639                                                                 - - ATTC                 - #                  - #                  - #                643                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:20:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 188 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                              - - Leu Asn Arg Thr Asp Pro Asn Ile Lys Leu Va - #l Leu Asp Asp Cys Trp        1               5 - #                 10 - #                 15              - - Ala Thr Ser Thr Met Asp Pro Ala Ser Leu Pr - #o Gln Trp Asn Ile Ile                   20     - #             25     - #             30                  - - Val Asp Gly Cys Glu Tyr Asn Leu Asp Asn Hi - #s Arg Thr Thr Phe His               35         - #         40         - #         45                      - - Pro Val Gly Ser Ser Val Ala Tyr Pro Asn Hi - #s Tyr Gln Arg Phe Ala           50             - #     55             - #     60                          - - Val Lys Thr Phe Ala Phe Val Ser Glu Asp Pr - #o Ala Phe Ser His Leu       65                 - # 70                 - # 75                 - # 80       - - Val Tyr Phe His Cys Ser Ala Leu Ile Cys As - #p Gln Leu Ser Ser Asn                       85 - #                 90 - #                 95              - - Phe Pro Leu Cys Ser Ala Ser Cys Leu Val Se - #r Ser Arg Ser Arg Arg                  100      - #           105      - #           110                  - - Ala Thr Gly Ala Thr Glu Glu Glu Lys Met Il - #e Val Ser Leu Pro Gly              115          - #       120          - #       125                      - - Pro Ile Leu Leu Leu Ser Asp Gly Ser Ser Ph - #e Arg Asp Ala Val Asp          130              - #   135              - #   140                          - - Ser Lys Gly His Gly Thr Ser Gly Tyr Ala Al - #a Phe Lys Thr Met Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ala Val Val Ala Leu Ala Gly Val Val Ala Th - #r Leu Ser Leu Ile        Ser                                                                                             165  - #               170  - #               175             - - Tyr Leu Arg Lys Lys Arg Ile Thr Val Leu As - #n His                                  180      - #           185                                         - -  - - (2) INFORMATION FOR SEQ ID NO:21:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1029 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bos tauru - #s                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..976                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                              - - G AAT TCT GTA CAC TTG GCC TTC AGG AAT GAC - #AGC GAA TGT AAA CCT             46                                                                          Asn Ser Val His Leu Ala Phe Arg Asn A - #sp Ser Glu Cys Lys Pro                 1              - # 5                 - # 10                 - # 15         - - GTG ATG GCA ACA CAC ACT TTT GTT CTG TTC CG - #G TTT CCA TTT ACT ACT           94                                                                       Val Met Ala Thr His Thr Phe Val Leu Phe Ar - #g Phe Pro Phe Thr Thr                            20 - #                 25 - #                 30              - - TGT GGT ACT ACA AAA CAG ATC ACT GGA AAG CA - #A GCG GTA TAT GAA AAT          142                                                                       Cys Gly Thr Thr Lys Gln Ile Thr Gly Lys Gl - #n Ala Val Tyr Glu Asn                        35     - #             40     - #             45                  - - GAG CTG GTT GCA GCT CGG GAT GTG AGA ACT TG - #G AGC CGT GGT TCT ATT          190                                                                       Glu Leu Val Ala Ala Arg Asp Val Arg Thr Tr - #p Ser Arg Gly Ser Ile                    50         - #         55         - #         60                      - - ACC CGA GAC AGT ACC TTC AGG CTC CAA GTC AG - #T TGT AGC TAC TCT GCA          238                                                                       Thr Arg Asp Ser Thr Phe Arg Leu Gln Val Se - #r Cys Ser Tyr Ser Ala                65             - #     70             - #     75                          - - AGT AGC AGT GCT CTC CCA GTT AAT GTC CAA GT - #T CTT ACT CTC CCA CCA          286                                                                       Ser Ser Ser Ala Leu Pro Val Asn Val Gln Va - #l Leu Thr Leu Pro Pro            80                 - # 85                 - # 90                 - # 95       - - CCC CTT CCT GAG ACC CTG CCT GGA AAC CTC AC - #T CTG GAA CTT AAG ATT          334                                                                       Pro Leu Pro Glu Thr Leu Pro Gly Asn Leu Th - #r Leu Glu Leu Lys Ile                           100  - #               105  - #               110              - - GCC AAA GAT AAA CCG TAT CGC TCC TAC TAC AC - #G GCT AGT GAC TAC CCA          382                                                                       Ala Lys Asp Lys Pro Tyr Arg Ser Tyr Tyr Th - #r Ala Ser Asp Tyr Pro                       115      - #           120      - #           125                  - - GTG GTG AAG TTA CTT CGG GAT CCC ATC TAC GT - #G GAA GTC TCC ATC CAT          430                                                                       Val Val Lys Leu Leu Arg Asp Pro Ile Tyr Va - #l Glu Val Ser Ile His                   130          - #       135          - #       140                      - - CAG AGA ACA GAC CCC AGT CTC GAG CTG CGC CT - #G GAC CAG TGT TGG GCG          478                                                                       Gln Arg Thr Asp Pro Ser Leu Glu Leu Arg Le - #u Asp Gln Cys Trp Ala               145              - #   150              - #   155                          - - ACA CCT GGT GCA GAT GCC CTG CTC CAG CCC CA - #G TGG CCC TTG CTT GTG          526                                                                       Thr Pro Gly Ala Asp Ala Leu Leu Gln Pro Gl - #n Trp Pro Leu Leu Val           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - AAT GGG TGC CCC TAC ACA GGA GAC AAC TAT CA - #G ACA AAA CTG ATC        CCT      574                                                                    Asn Gly Cys Pro Tyr Thr Gly Asp Asn Tyr Gl - #n Thr Lys Leu Ile Pro                          180  - #               185  - #               190              - - GTC TGG GAA GCC TCA GAC CTG CCG TTT CCT TC - #T CAC TAC CAG CGC TTC          622                                                                       Val Trp Glu Ala Ser Asp Leu Pro Phe Pro Se - #r His Tyr Gln Arg Phe                       195      - #           200      - #           205                  - - AGC ATT TCC ACC TTC AGC TTT GTG GAC TCA GT - #G GCA AAG CGG GCC CTC          670                                                                       Ser Ile Ser Thr Phe Ser Phe Val Asp Ser Va - #l Ala Lys Arg Ala Leu                   210          - #       215          - #       220                      - - AAG GGA CCG GTG TAT CTG CAC TGC AGT GCA TC - #G GTC TGC CAG CCT GCC          718                                                                       Lys Gly Pro Val Tyr Leu His Cys Ser Ala Se - #r Val Cys Gln Pro Ala               225              - #   230              - #   235                          - - GGG ACA CCA TCC TGT GTG ACA CTC TGT CCT GC - #C AGA CGA AGA AGA AGC          766                                                                       Gly Thr Pro Ser Cys Val Thr Leu Cys Pro Al - #a Arg Arg Arg Arg Ser           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - TCT GAC ATC CAT TTT CAG AAC AAA ACG GCT AG - #C ATT TCT AGC AAG        GGT      814                                                                    Ser Asp Ile His Phe Gln Asn Lys Thr Ala Se - #r Ile Ser Ser Lys Gly                          260  - #               265  - #               270              - - CCC TTG ATT CTA CTC CAA GCC ATT CAA GAC TC - #T TCA GAA AAG CTC CAC          862                                                                       Pro Leu Ile Leu Leu Gln Ala Ile Gln Asp Se - #r Ser Glu Lys Leu His                       275      - #           280      - #           285                  - - AAA TAC TCA AGG TCT CCT GTA GAC TCT CAA GC - #T TTG TGG GTG GCT GGC          910                                                                       Lys Tyr Ser Arg Ser Pro Val Asp Ser Gln Al - #a Leu Trp Val Ala Gly                   290          - #       295          - #       300                      - - CTA TCT GGA ATC TTA ATC GTT GGA GCC TTG TT - #C ATG TCC TAC CTG GCC          958                                                                       Leu Ser Gly Ile Leu Ile Val Gly Ala Leu Ph - #e Met Ser Tyr Leu Ala               305              - #   310              - #   315                          - - ATT AGG AAA TGG AGA TGAGTTGCTC AGCCCAAATG TGTTAATAA - #A ACCAGATTGC         1013                                                                       Ile Arg Lys Trp Arg                                                           320                                                                            - - AGCCGGCCGC GAATTC             - #                  - #                      - #  1029                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:22:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 324 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                              - - Asn Ser Val His Leu Ala Phe Arg Asn Asp Se - #r Glu Cys Lys Pro Val        1               5 - #                 10 - #                 15              - - Met Ala Thr His Thr Phe Val Leu Phe Arg Ph - #e Pro Phe Thr Thr Cys                   20     - #             25     - #             30                  - - Gly Thr Thr Lys Gln Ile Thr Gly Lys Gln Al - #a Val Tyr Glu Asn Glu               35         - #         40         - #         45                      - - Leu Val Ala Ala Arg Asp Val Arg Thr Trp Se - #r Arg Gly Ser Ile Thr           50             - #     55             - #     60                          - - Arg Asp Ser Thr Phe Arg Leu Gln Val Ser Cy - #s Ser Tyr Ser Ala Ser       65                 - # 70                 - # 75                 - # 80       - - Ser Ser Ala Leu Pro Val Asn Val Gln Val Le - #u Thr Leu Pro Pro Pro                       85 - #                 90 - #                 95              - - Leu Pro Glu Thr Leu Pro Gly Asn Leu Thr Le - #u Glu Leu Lys Ile Ala                  100      - #           105      - #           110                  - - Lys Asp Lys Pro Tyr Arg Ser Tyr Tyr Thr Al - #a Ser Asp Tyr Pro Val              115          - #       120          - #       125                      - - Val Lys Leu Leu Arg Asp Pro Ile Tyr Val Gl - #u Val Ser Ile His Gln          130              - #   135              - #   140                          - - Arg Thr Asp Pro Ser Leu Glu Leu Arg Leu As - #p Gln Cys Trp Ala Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Pro Gly Ala Asp Ala Leu Leu Gln Pro Gln Tr - #p Pro Leu Leu Val        Asn                                                                                             165  - #               170  - #               175             - - Gly Cys Pro Tyr Thr Gly Asp Asn Tyr Gln Th - #r Lys Leu Ile Pro Val                  180      - #           185      - #           190                  - - Trp Glu Ala Ser Asp Leu Pro Phe Pro Ser Hi - #s Tyr Gln Arg Phe Ser              195          - #       200          - #       205                      - - Ile Ser Thr Phe Ser Phe Val Asp Ser Val Al - #a Lys Arg Ala Leu Lys          210              - #   215              - #   220                          - - Gly Pro Val Tyr Leu His Cys Ser Ala Ser Va - #l Cys Gln Pro Ala Gly      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Thr Pro Ser Cys Val Thr Leu Cys Pro Ala Ar - #g Arg Arg Arg Ser        Ser                                                                                             245  - #               250  - #               255             - - Asp Ile His Phe Gln Asn Lys Thr Ala Ser Il - #e Ser Ser Lys Gly Pro                  260      - #           265      - #           270                  - - Leu Ile Leu Leu Gln Ala Ile Gln Asp Ser Se - #r Glu Lys Leu His Lys              275          - #       280          - #       285                      - - Tyr Ser Arg Ser Pro Val Asp Ser Gln Ala Le - #u Trp Val Ala Gly Leu          290              - #   295              - #   300                          - - Ser Gly Ile Leu Ile Val Gly Ala Leu Phe Me - #t Ser Tyr Leu Ala Ile      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Arg Lys Trp Arg                                                           - -  - - (2) INFORMATION FOR SEQ ID NO:23:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1457 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bos tauru - #s                                                  (D) DEVELOPMENTAL STAGE: - #Juvenile                                          (E) HAPLOTYPE: Diploidy                                                       (F) TISSUE TYPE: Ovary                                                        (G) CELL TYPE: Oocyte                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 149..1411                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                              - - CCCGGGCCTC CCTACTCTCA GGAAGGCACC CGCTCACCTC CTCAAGTTCT CG -            #ATCTCGGC     60                                                                 - - CGGGATGCTC TGAAGCTGGT TGCCGCCGAG GCTGAGGGTC TGCAGCGGCG CA -            #GTCCAGCA    120                                                                 - - GCGAGGTGGG AGTGGCTTCG TGGGCACC ATG GGG CCG TGC TCT - #AGG CTG TTC           172                                                                                         - #             Met Gly Pro - #Cys Ser Arg Leu Phe                            - #               1   - #            5                       - - GTC TGC TTT CTG CTC TGG GGA AGC ACA GAG CT - #C TGC AGC CCC CAG CCC          220                                                                       Val Cys Phe Leu Leu Trp Gly Ser Thr Glu Le - #u Cys Ser Pro Gln Pro                10             - #     15             - #     20                          - - TTC TGG GAT GAT GAA ACC GAG CGC TTC AGG CC - #A TCA AAG CCG CCC GCC          268                                                                       Phe Trp Asp Asp Glu Thr Glu Arg Phe Arg Pr - #o Ser Lys Pro Pro Ala            25                 - # 30                 - # 35                 - # 40       - - GTG ATG GTG GAG TGT CAG GAG GCC CAG CTG GT - #G GTC ACA GTC GAC AAA          316                                                                       Val Met Val Glu Cys Gln Glu Ala Gln Leu Va - #l Val Thr Val Asp Lys                            45 - #                 50 - #                 55              - - GAC CTT TTC GGC ACA GGG AAG CTC ATC CGG CC - #T GCG GAC CTC ACC CTG          364                                                                       Asp Leu Phe Gly Thr Gly Lys Leu Ile Arg Pr - #o Ala Asp Leu Thr Leu                        60     - #             65     - #             70                  - - GGC CCC GAC AAC TGT GAG CCG CTG GCC TCC GC - #G GAC ACG GAT GGC GTG          412                                                                       Gly Pro Asp Asn Cys Glu Pro Leu Ala Ser Al - #a Asp Thr Asp Gly Val                    75         - #         80         - #         85                      - - GTT AGG TTT GCG GTC GGG CTG CAC GAG TGT GG - #C AAC ATC TTG CAG GTG          460                                                                       Val Arg Phe Ala Val Gly Leu His Glu Cys Gl - #y Asn Ile Leu Gln Val                90             - #     95             - #    100                          - - ACC GAC AAT GCC CTG GTG TAC AGC ACC TTC CT - #G CTC CAC AAC CCC CGC          508                                                                       Thr Asp Asn Ala Leu Val Tyr Ser Thr Phe Le - #u Leu His Asn Pro Arg           105                 1 - #10                 1 - #15                 1 -      #20                                                                              - - CCT GCA GGA AAC CTG TCC ATC CTG AGG ACT AA - #C CGC GCA GAG GTC        CCC      556                                                                    Pro Ala Gly Asn Leu Ser Ile Leu Arg Thr As - #n Arg Ala Glu Val Pro                          125  - #               130  - #               135              - - ATC GAG TGC CAC TAC CCC AGG CAG GGC AAT GT - #G AGT AGC TGG GCC ATC          604                                                                       Ile Glu Cys His Tyr Pro Arg Gln Gly Asn Va - #l Ser Ser Trp Ala Ile                       140      - #           145      - #           150                  - - CAG CCC ACC TGG GTG CCA TTC AGG ACC ACA GT - #G TTC TCG GAG GAG AAG          652                                                                       Gln Pro Thr Trp Val Pro Phe Arg Thr Thr Va - #l Phe Ser Glu Glu Lys                   155          - #       160          - #       165                      - - CTG GTT TTC TCT CTG CGC CTG ATG GAG GAG AA - #C TGG AGC GCC GAG AAG          700                                                                       Leu Val Phe Ser Leu Arg Leu Met Glu Glu As - #n Trp Ser Ala Glu Lys               170              - #   175              - #   180                          - - ATG ACG CCC ACC TTC CAG CTG GGA GAC AGA GC - #C CAC CTC CAG GCC CAA          748                                                                       Met Thr Pro Thr Phe Gln Leu Gly Asp Arg Al - #a His Leu Gln Ala Gln           185                 1 - #90                 1 - #95                 2 -      #00                                                                              - - GTG CAC ACT GGC AGC CAC GTG CCC CTG CGG CT - #G TTC GTG GAC CAC        TGC      796                                                                    Val His Thr Gly Ser His Val Pro Leu Arg Le - #u Phe Val Asp His Cys                          205  - #               210  - #               215              - - GTG GCC AGC CTG ACG CCA GAC TGG AGC ACC TC - #C CCT TAC CAC ACC ATC          844                                                                       Val Ala Ser Leu Thr Pro Asp Trp Ser Thr Se - #r Pro Tyr His Thr Ile                       220      - #           225      - #           230                  - - GTG GAC TTC CAT GGT TGT CTC GTC GAT GGT CT - #C ACC GAT GCC TCC TCT          892                                                                       Val Asp Phe His Gly Cys Leu Val Asp Gly Le - #u Thr Asp Ala Ser Ser                   235          - #       240          - #       245                      - - GCT TTC AAA GCA CCC AGA CCC AGA CCG GAG AT - #C CTC CAG TTC ACA GTG          940                                                                       Ala Phe Lys Ala Pro Arg Pro Arg Pro Glu Il - #e Leu Gln Phe Thr Val               250              - #   255              - #   260                          - - GAT GTG TTC CGT TTT GCT AAT GAC TCC AGA AA - #C ATG ATA TAT ATC ACC          988                                                                       Asp Val Phe Arg Phe Ala Asn Asp Ser Arg As - #n Met Ile Tyr Ile Thr           265                 2 - #70                 2 - #75                 2 -      #80                                                                              - - TGC CAC CTG AAG GTC ACT CCG GTT GAC CGA GT - #C CCG GAC CAA CTA        AAC     1036                                                                    Cys His Leu Lys Val Thr Pro Val Asp Arg Va - #l Pro Asp Gln Leu Asn                          285  - #               290  - #               295              - - AAA GCC TGT TCC TTC AGC AAG TCC TCC AAC AG - #G TGG TCC CCG GTT GAA         1084                                                                       Lys Ala Cys Ser Phe Ser Lys Ser Ser Asn Ar - #g Trp Ser Pro Val Glu                       300      - #           305      - #           310                  - - GGC CCC ACT GAC ATC TGT CGA TGC TGT AGC AA - #G GGG CGC TGT GGC ATT         1132                                                                       Gly Pro Thr Asp Ile Cys Arg Cys Cys Ser Ly - #s Gly Arg Cys Gly Ile                   315          - #       320          - #       325                      - - TCA GGC CGT TCC ATG AGG CTG TCC CAC CGG GA - #G GGC AGG CCT GTT CCC         1180                                                                       Ser Gly Arg Ser Met Arg Leu Ser His Arg Gl - #u Gly Arg Pro Val Pro               330              - #   335              - #   340                          - - CGA AGT CGC AGG CAC GTG ACG GAG GAA GCA GA - #T GTC ACC GTG GGG CCG         1228                                                                       Arg Ser Arg Arg His Val Thr Glu Glu Ala As - #p Val Thr Val Gly Pro           345                 3 - #50                 3 - #55                 3 -      #60                                                                              - - TTG ATC TTC CTG AGG AAG ATG AAT GAC CGT GG - #C GTG GAA GGG CCC        ACC     1276                                                                    Leu Ile Phe Leu Arg Lys Met Asn Asp Arg Gl - #y Val Glu Gly Pro Thr                          365  - #               370  - #               375              - - TCC TCT CCC CCT CTG GTG ATG CTG GGC TTA GG - #C CTG GCT ACT GTG ATG         1324                                                                       Ser Ser Pro Pro Leu Val Met Leu Gly Leu Gl - #y Leu Ala Thr Val Met                       380      - #           385      - #           390                  - - ACC TTG ACT CTG GCT GCC ATT GTC CTG GGT CT - #C ACT GGG AGG CTT CGG         1372                                                                       Thr Leu Thr Leu Ala Ala Ile Val Leu Gly Le - #u Thr Gly Arg Leu Arg                   395          - #       400          - #       405                      - - GCT GCT TCT CAC CCC GTG TGC CCT GTG TCT GC - #T TCC CAA TAAAAGAAGA          1421                                                                       Ala Ala Ser His Pro Val Cys Pro Val Ser Al - #a Ser Gln                           410              - #   415              - #   420                          - - AAGTGAAAAA AAAAAAAAAA AAGCGGCCGC GAATTC      - #                  -     #     1457                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:24:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 421 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                              - - Met Gly Pro Cys Ser Arg Leu Phe Val Cys Ph - #e Leu Leu Trp Gly Ser        1               5 - #                 10 - #                 15              - - Thr Glu Leu Cys Ser Pro Gln Pro Phe Trp As - #p Asp Glu Thr Glu Arg                   20     - #             25     - #             30                  - - Phe Arg Pro Ser Lys Pro Pro Ala Val Met Va - #l Glu Cys Gln Glu Ala               35         - #         40         - #         45                      - - Gln Leu Val Val Thr Val Asp Lys Asp Leu Ph - #e Gly Thr Gly Lys Leu           50             - #     55             - #     60                          - - Ile Arg Pro Ala Asp Leu Thr Leu Gly Pro As - #p Asn Cys Glu Pro Leu       65                 - # 70                 - # 75                 - # 80       - - Ala Ser Ala Asp Thr Asp Gly Val Val Arg Ph - #e Ala Val Gly Leu His                       85 - #                 90 - #                 95              - - Glu Cys Gly Asn Ile Leu Gln Val Thr Asp As - #n Ala Leu Val Tyr Ser                  100      - #           105      - #           110                  - - Thr Phe Leu Leu His Asn Pro Arg Pro Ala Gl - #y Asn Leu Ser Ile Leu              115          - #       120          - #       125                      - - Arg Thr Asn Arg Ala Glu Val Pro Ile Glu Cy - #s His Tyr Pro Arg Gln          130              - #   135              - #   140                          - - Gly Asn Val Ser Ser Trp Ala Ile Gln Pro Th - #r Trp Val Pro Phe Arg      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Thr Thr Val Phe Ser Glu Glu Lys Leu Val Ph - #e Ser Leu Arg Leu        Met                                                                                             165  - #               170  - #               175             - - Glu Glu Asn Trp Ser Ala Glu Lys Met Thr Pr - #o Thr Phe Gln Leu Gly                  180      - #           185      - #           190                  - - Asp Arg Ala His Leu Gln Ala Gln Val His Th - #r Gly Ser His Val Pro              195          - #       200          - #       205                      - - Leu Arg Leu Phe Val Asp His Cys Val Ala Se - #r Leu Thr Pro Asp Trp          210              - #   215              - #   220                          - - Ser Thr Ser Pro Tyr His Thr Ile Val Asp Ph - #e His Gly Cys Leu Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asp Gly Leu Thr Asp Ala Ser Ser Ala Phe Ly - #s Ala Pro Arg Pro        Arg                                                                                             245  - #               250  - #               255             - - Pro Glu Ile Leu Gln Phe Thr Val Asp Val Ph - #e Arg Phe Ala Asn Asp                  260      - #           265      - #           270                  - - Ser Arg Asn Met Ile Tyr Ile Thr Cys His Le - #u Lys Val Thr Pro Val              275          - #       280          - #       285                      - - Asp Arg Val Pro Asp Gln Leu Asn Lys Ala Cy - #s Ser Phe Ser Lys Ser          290              - #   295              - #   300                          - - Ser Asn Arg Trp Ser Pro Val Glu Gly Pro Th - #r Asp Ile Cys Arg Cys      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Cys Ser Lys Gly Arg Cys Gly Ile Ser Gly Ar - #g Ser Met Arg Leu        Ser                                                                                             325  - #               330  - #               335             - - His Arg Glu Gly Arg Pro Val Pro Arg Ser Ar - #g Arg His Val Thr Glu                  340      - #           345      - #           350                  - - Glu Ala Asp Val Thr Val Gly Pro Leu Ile Ph - #e Leu Arg Lys Met Asn              355          - #       360          - #       365                      - - Asp Arg Gly Val Glu Gly Pro Thr Ser Ser Pr - #o Pro Leu Val Met Leu          370              - #   375              - #   380                          - - Gly Leu Gly Leu Ala Thr Val Met Thr Leu Th - #r Leu Ala Ala Ile Val      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Leu Gly Leu Thr Gly Arg Leu Arg Ala Ala Se - #r His Pro Val Cys        Pro                                                                                             405  - #               410  - #               415             - - Val Ser Ala Ser Gln                                                                  420                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:25:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 125 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                              - - AGTTCGTGCT TATCTGAACA TGTCTTGAGG GATTAGTATG TGTGCTCATT TG -             #GGTTCTTT     60                                                                 - - CCGCTGTATG CTAGGCGTAT CTAGATGCAT TAGCTTGTTA ACACCTCATG TG -            #GAGTAAAA    120                                                                 - - GATGT                 - #                  - #                  -      #           125                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:26:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 111 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                              - - CAGGCGTAGG CGTGGACTGA AGTTCAAAGC CATGCGCCCG TTCTGATAGC AT -            #ACGTTTGA     60                                                                 - - AATGTCATTG TAGTTGCATG GCTGTATAAG CCAGTCTCAT AGATAAGGGA A - #                111                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:27:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 96 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                              - - GCGGTCGGTC ATGTGATGCT GCGTATAGTA CGATTTTGAA TGCATTATGC GA -             #AATTATTC     60                                                                 - - TAACGACCCG CGATATGGAG GTTGGATTAA GTTACA      - #                       - #       96                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:28:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 19 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                              - - ATGGARAGRT GYCAMGARG             - #                  - #                      - # 19                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:29:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                              - - GATCTAAGGA AGATCTATGG ATCC          - #                  - #                    24                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:30:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                              - - GATCTAAGGA GGTTGTATGG ATCC          - #                  - #                    24                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:31:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 55 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                              - - GATCTATGAC CATGATTACG GATTCGCGTA GCCGTCGTCC TGCAGCGTCG CG - #ACT              55                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:32:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 57 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                              - - GGGAAAACCC GGGCGTTACC CAACTTAATC GATTAGCAGC ACATCCCCCT TC - #GCCAG            57                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:33:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 54 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                              - - TTTTCCCAGT CGCGCTGCAG AACGACGGCT AGCGAATCCG TAATCATGGT CA - #TA               54                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:34:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 52 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                              - - CTGGCCAAAG GGGGATGTGG CTGCTAATCG ATTAAGTTGG GTAACGCCCG GG - #                 52                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:35:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 220 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                              - - GATCTATGAC CATGATTACG GATTCGCTAG CCGTCGTTCT GCAGCGTCGC GA -             #CTGGGAAA     60                                                                 - - ATACTGGTAC TAATGCCTAA GCGATCGGCA GCAAGACGTC GGAGCGCTGAC C -            #CTTTACCC    120                                                                 - - GGGCGTTACC CAACTTAATC GATTAGCAGC ACATCCCCCT TTCGCCAGTGG G -            #CCCGCAAT    180                                                                 - - CCCTTGAATT AGCAAATCGT CGTGTAGGGG GAAAGCGGTC     - #                      - #   220                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:36:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                              - - GCGAAGCTTC CGACACCATC GAACGGCGC         - #                  - #                29                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:37:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                              - - GCGCACAATG TGCCTAATGA GTGAGCTAAC         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:38:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                              - - CGCGGATCCG GACGAAGGCC AGCGCTTG         - #                  - #                 28                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:39:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 58 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                              - - GCGGTCGACT CATTAATGAT GATGATGATG ATGCGGGCTC GAGGTGGACC CT - #TCCACC           58                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:40:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1701 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1698                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                              - - ATG TGG CTG CTG CGG TGC GTT TTG CTG TGT GT - #T TCA TTA TCT CTT GCT           48                                                                       Met Trp Leu Leu Arg Cys Val Leu Leu Cys Va - #l Ser Leu Ser Leu Ala             1               5 - #                 10 - #                 15              - - GTG AGT GGC CAG CAT AAG CCT GAG GCA CCA GA - #T TAT TCC AGT GTG CTC           96                                                                       Val Ser Gly Gln His Lys Pro Glu Ala Pro As - #p Tyr Ser Ser Val Leu                        20     - #             25     - #             30                  - - CAC TGT GGG CCG TGG AGC TTC CAG TTT GCT GT - #A AAC CTC AAC CAG GAG          144                                                                       His Cys Gly Pro Trp Ser Phe Gln Phe Ala Va - #l Asn Leu Asn Gln Glu                    35         - #         40         - #         45                      - - GCA ACG TCT CCT CCT GTA CTA ATA GCT TGG GA - #C AAC CAA GGG CTG CTG          192                                                                       Ala Thr Ser Pro Pro Val Leu Ile Ala Trp As - #p Asn Gln Gly Leu Leu                50             - #     55             - #     60                          - - CAC GAG CTG CAG AAT GAC TCC GAC TGT GGC AC - #C TGG ATA AGA AAA GGT          240                                                                       His Glu Leu Gln Asn Asp Ser Asp Cys Gly Th - #r Trp Ile Arg Lys Gly            65                 - # 70                 - # 75                 - # 80       - - CCA GGC AGC TCC GTG GTG TTG GAG GCA ACC TA - #T AGC AGC TGC TAT GTC          288                                                                       Pro Gly Ser Ser Val Val Leu Glu Ala Thr Ty - #r Ser Ser Cys Tyr Val                            85 - #                 90 - #                 95              - - ACT GAG TGG GTG AGT ATG ACC CAA TGG CCA GG - #G AGA CTG TGT GAA GCG          336                                                                       Thr Glu Trp Val Ser Met Thr Gln Trp Pro Gl - #y Arg Leu Cys Glu Ala                       100      - #           105      - #           110                  - - CCT CAT GCT ACC ATC CAG GCT GAC CCC CAA GG - #C CTG TCT CTC CAG GAC          384                                                                       Pro His Ala Thr Ile Gln Ala Asp Pro Gln Gl - #y Leu Ser Leu Gln Asp                   115          - #       120          - #       125                      - - TCC CAC TAC ATC ATG CCA GTT GGA GTT GAA GG - #A GCA GGC GCG GCT GAA          432                                                                       Ser His Tyr Ile Met Pro Val Gly Val Glu Gl - #y Ala Gly Ala Ala Glu               130              - #   135              - #   140                          - - CAC AAG GTG GTT ACA GAG AGG AAG CTG CTC AA - #G TGT CCT ATG GAT CTT          480                                                                       His Lys Val Val Thr Glu Arg Lys Leu Leu Ly - #s Cys Pro Met Asp Leu           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - CTA GAT GCT CCA GAT ACT GAC TGG TGT GAC TC - #C ATC CCA GCA CGG        GAC      528                                                                    Leu Asp Ala Pro Asp Thr Asp Trp Cys Asp Se - #r Ile Pro Ala Arg Asp                          165  - #               170  - #               175              - - AGA CTG CCA TGT GCA CCT TCA CCC ATC TCT CG - #A GGA GAC TGT GAA GGG          576                                                                       Arg Leu Pro Cys Ala Pro Ser Pro Ile Ser Ar - #g Gly Asp Cys Glu Gly                       180      - #           185      - #           190                  - - CTA GGC TGT TGT TAT AGC TCT GAA GAG GTG AA - #T TCC TGC TAC TAT GGA          624                                                                       Leu Gly Cys Cys Tyr Ser Ser Glu Glu Val As - #n Ser Cys Tyr Tyr Gly                   195          - #       200          - #       205                      - - AAC ACT GTG ACC TTG CAT TGT ACC CGA GAG GG - #C CAT TTC TCT ATT GCT          672                                                                       Asn Thr Val Thr Leu His Cys Thr Arg Glu Gl - #y His Phe Ser Ile Ala               210              - #   215              - #   220                          - - GTG TCT CGG AAC GTG ACC TCG CCA CCA CTG CT - #C TTG GAT TCT GTG CGC          720                                                                       Val Ser Arg Asn Val Thr Ser Pro Pro Leu Le - #u Leu Asp Ser Val Arg           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - TTG GCC CTT AGG AAT GAC AGT GCG TGT AAC CC - #T GTG ATG GCA ACA        CAA      768                                                                    Leu Ala Leu Arg Asn Asp Ser Ala Cys Asn Pr - #o Val Met Ala Thr Gln                          245  - #               250  - #               255              - - GCT TTT GTT CTG TTC CAG TTT CCA TTT ACT TC - #C TGT GGC ACC ACA AGA          816                                                                       Ala Phe Val Leu Phe Gln Phe Pro Phe Thr Se - #r Cys Gly Thr Thr Arg                       260      - #           265      - #           270                  - - CAG ATC ACT GGA GAC CGA GCA GTA TAT GAA AA - #T GAA CTG GTG GCA ACT          864                                                                       Gln Ile Thr Gly Asp Arg Ala Val Tyr Glu As - #n Glu Leu Val Ala Thr                   275          - #       280          - #       285                      - - AGG GAT GTG AAA AAT GGG AGC CGT GGC TCT GT - #C ACT CGT GAC AGC ATC          912                                                                       Arg Asp Val Lys Asn Gly Ser Arg Gly Ser Va - #l Thr Arg Asp Ser Ile               290              - #   295              - #   300                          - - TTC AGG CTC CAT GTC AGC TGC AGC TAC TCA GT - #A AGT AGC AAC TCT CTC          960                                                                       Phe Arg Leu His Val Ser Cys Ser Tyr Ser Va - #l Ser Ser Asn Ser Leu           305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - CCA ATC AAT GTC CAG GTT TTC ACT CTC CCA CC - #A CCC TTT CCT GAG        ACC     1008                                                                    Pro Ile Asn Val Gln Val Phe Thr Leu Pro Pr - #o Pro Phe Pro Glu Thr                          325  - #               330  - #               335              - - CAG CCT GGA CCC CTC ACT CTG GAA CTT CAG AT - #T GCC AAA GAT AAA AAC         1056                                                                       Gln Pro Gly Pro Leu Thr Leu Glu Leu Gln Il - #e Ala Lys Asp Lys Asn                       340      - #           345      - #           350                  - - TAT GGC TCT TAC TAC GGT GTT GGT GAC TAC CC - #A GTG GTG AAG TTG CTT         1104                                                                       Tyr Gly Ser Tyr Tyr Gly Val Gly Asp Tyr Pr - #o Val Val Lys Leu Leu                   355          - #       360          - #       365                      - - CGG GAT CCC ATT TAC GTG GAG GTC TCC ATC CT - #T CAC AGA ACA GAC CCC         1152                                                                       Arg Asp Pro Ile Tyr Val Glu Val Ser Ile Le - #u His Arg Thr Asp Pro               370              - #   375              - #   380                          - - TAC CTG GGG CTG CTC CTA CAA CAG TGT TGG GC - #A ACA CCC AGC ACT GAC         1200                                                                       Tyr Leu Gly Leu Leu Leu Gln Gln Cys Trp Al - #a Thr Pro Ser Thr Asp           385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - CCC CTG AGT CAG CCA CAG TGG CCC ATC CTG GT - #A AAG GGC TGC CCC        TAC     1248                                                                    Pro Leu Ser Gln Pro Gln Trp Pro Ile Leu Va - #l Lys Gly Cys Pro Tyr                          405  - #               410  - #               415              - - ATT GGA GAC AAC TAT CAG ACC CAG CTG ATC CC - #T GTC CAG AAA GCC TTG         1296                                                                       Ile Gly Asp Asn Tyr Gln Thr Gln Leu Ile Pr - #o Val Gln Lys Ala Leu                       420      - #           425      - #           430                  - - GAT CTT CCA TTT CCC TCT CAC CAC CAG CGC TT - #C AGC ATC TTC ACC TTC         1344                                                                       Asp Leu Pro Phe Pro Ser His His Gln Arg Ph - #e Ser Ile Phe Thr Phe                   435          - #       440          - #       445                      - - AGC TTT GTG AAC CCT ACA GTG GAG AAA CAG GC - #C CTC AGG GGA CCG GTG         1392                                                                       Ser Phe Val Asn Pro Thr Val Glu Lys Gln Al - #a Leu Arg Gly Pro Val               450              - #   455              - #   460                          - - CAT CTG CAC TGC AGC GTG TCA GTC TGC CAG CC - #T GCT GAG ACA CCA TCC         1440                                                                       His Leu His Cys Ser Val Ser Val Cys Gln Pr - #o Ala Glu Thr Pro Ser           465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - TGT GTG GTG ACC TGT CCT GAT CTC AGT CGA AG - #A AGA AAT TTT GAC        AAC     1488                                                                    Cys Val Val Thr Cys Pro Asp Leu Ser Arg Ar - #g Arg Asn Phe Asp Asn                          485  - #               490  - #               495              - - AGT TCT CAG AAC ACT ACT GCT AGT GTT TCT AG - #C AAA GGC CCC ATG ATT         1536                                                                       Ser Ser Gln Asn Thr Thr Ala Ser Val Ser Se - #r Lys Gly Pro Met Ile                       500      - #           505      - #           510                  - - CTA CTC CAA GCC ACT AAG GAC CCT CCA GAA AA - #G CTC CGT GTT CCT GTA         1584                                                                       Leu Leu Gln Ala Thr Lys Asp Pro Pro Glu Ly - #s Leu Arg Val Pro Val                   515          - #       520          - #       525                      - - GAC TCG AAA GTT CTG TGG GTG GCA GGC CTT TC - #T GGG ACC TTA ATC CTT         1632                                                                       Asp Ser Lys Val Leu Trp Val Ala Gly Leu Se - #r Gly Thr Leu Ile Leu               530              - #   535              - #   540                          - - GGA GCC TTG TTA GTA TCC TAC TTG GCT GTC AA - #G AAA CAG AAG AGT TGC         1680                                                                       Gly Ala Leu Leu Val Ser Tyr Leu Ala Val Ly - #s Lys Gln Lys Ser Cys           545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - CCA GAC CAA ATG TGT CAA TAA       - #                  - #                    1701                                                                    Pro Asp Gln Met Cys Gln                                                                       565                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:41:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 566 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                              - - Met Trp Leu Leu Arg Cys Val Leu Leu Cys Va - #l Ser Leu Ser Leu Ala        1               5 - #                 10 - #                 15              - - Val Ser Gly Gln His Lys Pro Glu Ala Pro As - #p Tyr Ser Ser Val Leu                   20     - #             25     - #             30                  - - His Cys Gly Pro Trp Ser Phe Gln Phe Ala Va - #l Asn Leu Asn Gln Glu               35         - #         40         - #         45                      - - Ala Thr Ser Pro Pro Val Leu Ile Ala Trp As - #p Asn Gln Gly Leu Leu           50             - #     55             - #     60                          - - His Glu Leu Gln Asn Asp Ser Asp Cys Gly Th - #r Trp Ile Arg Lys Gly       65                 - # 70                 - # 75                 - # 80       - - Pro Gly Ser Ser Val Val Leu Glu Ala Thr Ty - #r Ser Ser Cys Tyr Val                       85 - #                 90 - #                 95              - - Thr Glu Trp Val Ser Met Thr Gln Trp Pro Gl - #y Arg Leu Cys Glu Ala                  100      - #           105      - #           110                  - - Pro His Ala Thr Ile Gln Ala Asp Pro Gln Gl - #y Leu Ser Leu Gln Asp              115          - #       120          - #       125                      - - Ser His Tyr Ile Met Pro Val Gly Val Glu Gl - #y Ala Gly Ala Ala Glu          130              - #   135              - #   140                          - - His Lys Val Val Thr Glu Arg Lys Leu Leu Ly - #s Cys Pro Met Asp Leu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Leu Asp Ala Pro Asp Thr Asp Trp Cys Asp Se - #r Ile Pro Ala Arg        Asp                                                                                             165  - #               170  - #               175             - - Arg Leu Pro Cys Ala Pro Ser Pro Ile Ser Ar - #g Gly Asp Cys Glu Gly                  180      - #           185      - #           190                  - - Leu Gly Cys Cys Tyr Ser Ser Glu Glu Val As - #n Ser Cys Tyr Tyr Gly              195          - #       200          - #       205                      - - Asn Thr Val Thr Leu His Cys Thr Arg Glu Gl - #y His Phe Ser Ile Ala          210              - #   215              - #   220                          - - Val Ser Arg Asn Val Thr Ser Pro Pro Leu Le - #u Leu Asp Ser Val Arg      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Leu Ala Leu Arg Asn Asp Ser Ala Cys Asn Pr - #o Val Met Ala Thr        Gln                                                                                             245  - #               250  - #               255             - - Ala Phe Val Leu Phe Gln Phe Pro Phe Thr Se - #r Cys Gly Thr Thr Arg                  260      - #           265      - #           270                  - - Gln Ile Thr Gly Asp Arg Ala Val Tyr Glu As - #n Glu Leu Val Ala Thr              275          - #       280          - #       285                      - - Arg Asp Val Lys Asn Gly Ser Arg Gly Ser Va - #l Thr Arg Asp Ser Ile          290              - #   295              - #   300                          - - Phe Arg Leu His Val Ser Cys Ser Tyr Ser Va - #l Ser Ser Asn Ser Leu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Pro Ile Asn Val Gln Val Phe Thr Leu Pro Pr - #o Pro Phe Pro Glu        Thr                                                                                             325  - #               330  - #               335             - - Gln Pro Gly Pro Leu Thr Leu Glu Leu Gln Il - #e Ala Lys Asp Lys Asn                  340      - #           345      - #           350                  - - Tyr Gly Ser Tyr Tyr Gly Val Gly Asp Tyr Pr - #o Val Val Lys Leu Leu              355          - #       360          - #       365                      - - Arg Asp Pro Ile Tyr Val Glu Val Ser Ile Le - #u His Arg Thr Asp Pro          370              - #   375              - #   380                          - - Tyr Leu Gly Leu Leu Leu Gln Gln Cys Trp Al - #a Thr Pro Ser Thr Asp      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Pro Leu Ser Gln Pro Gln Trp Pro Ile Leu Va - #l Lys Gly Cys Pro        Tyr                                                                                             405  - #               410  - #               415             - - Ile Gly Asp Asn Tyr Gln Thr Gln Leu Ile Pr - #o Val Gln Lys Ala Leu                  420      - #           425      - #           430                  - - Asp Leu Pro Phe Pro Ser His His Gln Arg Ph - #e Ser Ile Phe Thr Phe              435          - #       440          - #       445                      - - Ser Phe Val Asn Pro Thr Val Glu Lys Gln Al - #a Leu Arg Gly Pro Val          450              - #   455              - #   460                          - - His Leu His Cys Ser Val Ser Val Cys Gln Pr - #o Ala Glu Thr Pro Ser      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Cys Val Val Thr Cys Pro Asp Leu Ser Arg Ar - #g Arg Asn Phe Asp        Asn                                                                                             485  - #               490  - #               495             - - Ser Ser Gln Asn Thr Thr Ala Ser Val Ser Se - #r Lys Gly Pro Met Ile                  500      - #           505      - #           510                  - - Leu Leu Gln Ala Thr Lys Asp Pro Pro Glu Ly - #s Leu Arg Val Pro Val              515          - #       520          - #       525                      - - Asp Ser Lys Val Leu Trp Val Ala Gly Leu Se - #r Gly Thr Leu Ile Leu          530              - #   535              - #   540                          - - Gly Ala Leu Leu Val Ser Tyr Leu Ala Val Ly - #s Lys Gln Lys Ser Cys      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Pro Asp Gln Met Cys Gln                                                                  565                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:42:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2266 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..2235                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                              - - ATG GCG TGC AGG CAG AGA GGA GGC TCT TGG AG - #T CCC TCA GGC TGG        TTC       48                                                                    Met Ala Cys Arg Gln Arg Gly Gly Ser Trp Se - #r Pro Ser Gly Trp Phe            1               5 - #                 10 - #                 15              - - AAT GCA GGC TGG AGC ACC TAC AGG TCG ATT TC - #T CTC TTC TTC GCC CTT           96                                                                       Asn Ala Gly Trp Ser Thr Tyr Arg Ser Ile Se - #r Leu Phe Phe Ala Leu                        20     - #             25     - #             30                  - - GTG ACT TCA GGG AAC TCC ATA GAT GTT TCT CA - #G TTG GTA AAT CCT GCC          144                                                                       Val Thr Ser Gly Asn Ser Ile Asp Val Ser Gl - #n Leu Val Asn Pro Ala                    35         - #         40         - #         45                      - - TTT CCA GGC ACT GTC ACT TGC GAT GAA AGG GA - #A ATA ACA GTG GAG TTC          192                                                                       Phe Pro Gly Thr Val Thr Cys Asp Glu Arg Gl - #u Ile Thr Val Glu Phe                50             - #     55             - #     60                          - - CCA AGC AGT CCT GGC ACC AAG AAA TGG CAT GC - #A TCT GTG GTG GAT CCT          240                                                                       Pro Ser Ser Pro Gly Thr Lys Lys Trp His Al - #a Ser Val Val Asp Pro            65                 - # 70                 - # 75                 - # 80       - - CTT GGT CTC GAC ATG CCG AAC TGC ACT TAC AT - #C CTG GAC CCA GAA AAG          288                                                                       Leu Gly Leu Asp Met Pro Asn Cys Thr Tyr Il - #e Leu Asp Pro Glu Lys                            85 - #                 90 - #                 95              - - CTC ACC CTG AGG GCT ACC TAT GAT AAC TGT AC - #C AGG AGA GTG CAT GGT          336                                                                       Leu Thr Leu Arg Ala Thr Tyr Asp Asn Cys Th - #r Arg Arg Val His Gly                       100      - #           105      - #           110                  - - GGA CAC CAG ATG ACC ATC AGA GTC ATG AAC AA - #C AGT GCT GCC TTA AGA          384                                                                       Gly His Gln Met Thr Ile Arg Val Met Asn As - #n Ser Ala Ala Leu Arg                   115          - #       120          - #       125                      - - CAC GGA GCT GTC ATG TAT CAG TTC TTC TGT CC - #A GCT ATG CAA GTA GAA          432                                                                       His Gly Ala Val Met Tyr Gln Phe Phe Cys Pr - #o Ala Met Gln Val Glu               130              - #   135              - #   140                          - - GAG ACC CAG GGG CTT TCA GCA TCT ACA ATC TG - #C CAG AAG GAT TTC ATG          480                                                                       Glu Thr Gln Gly Leu Ser Ala Ser Thr Ile Cy - #s Gln Lys Asp Phe Met           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - TCT TTT TCC TTG CCA CGG GTC TTC TCT GGC TT - #G GCT GAC GAC AGT        AAG      528                                                                    Ser Phe Ser Leu Pro Arg Val Phe Ser Gly Le - #u Ala Asp Asp Ser Lys                          165  - #               170  - #               175              - - GGG ACC AAA GTT CAG ATG GGA TGG AGC ATT GA - #G GTT GGT GAT GGT GCA          576                                                                       Gly Thr Lys Val Gln Met Gly Trp Ser Ile Gl - #u Val Gly Asp Gly Ala                       180      - #           185      - #           190                  - - AGA GCC AAA ACT CTG ACC CTG CCA GAG GCC AT - #G AAG GAA GGC TTC AGC          624                                                                       Arg Ala Lys Thr Leu Thr Leu Pro Glu Ala Me - #t Lys Glu Gly Phe Ser                   195          - #       200          - #       205                      - - CTC TTG ATT GAC AAC CAC AGG ATG ACC TTC CA - #T GTG CCA TTC AAT GCC          672                                                                       Leu Leu Ile Asp Asn His Arg Met Thr Phe Hi - #s Val Pro Phe Asn Ala               210              - #   215              - #   220                          - - ACT GGA GTG ACT CAC TAT GTG CAA GGT AAC AG - #T CAT CTC TAC ATG GTG          720                                                                       Thr Gly Val Thr His Tyr Val Gln Gly Asn Se - #r His Leu Tyr Met Val           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - TCT CTG AAG CTT ACA TTT ATA TCT CCT GGA CA - #G AAG GTG ATC TTC        TCT      768                                                                    Ser Leu Lys Leu Thr Phe Ile Ser Pro Gly Gl - #n Lys Val Ile Phe Ser                          245  - #               250  - #               255              - - TCA CAA GCT ATT TGT GCA CCA GAT CCT GTG AC - #C TGC AAT GCC ACA CAC          816                                                                       Ser Gln Ala Ile Cys Ala Pro Asp Pro Val Th - #r Cys Asn Ala Thr His                       260      - #           265      - #           270                  - - ATG ACT CTC ACC ATA CCA GAG TTT CCT GGG AA - #G CTT AAG TCT GTG AGC          864                                                                       Met Thr Leu Thr Ile Pro Glu Phe Pro Gly Ly - #s Leu Lys Ser Val Ser                   275          - #       280          - #       285                      - - TTT GAA AAC CAG AAC ATT GAT GTG AGC CAG CT - #G CAT GAC AAT GGA ATT          912                                                                       Phe Glu Asn Gln Asn Ile Asp Val Ser Gln Le - #u His Asp Asn Gly Ile               290              - #   295              - #   300                          - - GAT CTA GAA GCA ACA AAT GGC ATG AAA TTG CA - #T TTC AGC AAA ACT CTG          960                                                                       Asp Leu Glu Ala Thr Asn Gly Met Lys Leu Hi - #s Phe Ser Lys Thr Leu           305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - CTC AAA ACG AAA TTA TCT GAA AAA TGC CTA CT - #C CAT CAG TTC TAC        TTA     1008                                                                    Leu Lys Thr Lys Leu Ser Glu Lys Cys Leu Le - #u His Gln Phe Tyr Leu                          325  - #               330  - #               335              - - GCT TCA CTC AAG CTG ACC TTT CTC CTT CGG CC - #A GAG ACA GTA TCC ATG         1056                                                                       Ala Ser Leu Lys Leu Thr Phe Leu Leu Arg Pr - #o Glu Thr Val Ser Met                       340      - #           345      - #           350                  - - GTG ATC TAT CCT GAG TGT CTC TGT GAG TCA CC - #C GTT TCT ATA GTT ACA         1104                                                                       Val Ile Tyr Pro Glu Cys Leu Cys Glu Ser Pr - #o Val Ser Ile Val Thr                   355          - #       360          - #       365                      - - GGG GAG CTG TGC ACC CAG GAT GGG TTT ATG GA - #C GTC GAG GTC TAC AGC         1152                                                                       Gly Glu Leu Cys Thr Gln Asp Gly Phe Met As - #p Val Glu Val Tyr Ser               370              - #   375              - #   380                          - - TAC CAA ACA CAA CCA GCT CTT GAC CTG GGT AC - #T CTG AGG GTG GGA AAC         1200                                                                       Tyr Gln Thr Gln Pro Ala Leu Asp Leu Gly Th - #r Leu Arg Val Gly Asn           385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - TCA TCC TGC CAG CCT GTC TTT GAG GCT CAG TC - #T CAG GGG CTG GTA        CGG     1248                                                                    Ser Ser Cys Gln Pro Val Phe Glu Ala Gln Se - #r Gln Gly Leu Val Arg                          405  - #               410  - #               415              - - TTC CAC ATA CCC CTG AAT GGA TGT GGA ACG AG - #A TAT AAG TTC GAA GAT         1296                                                                       Phe His Ile Pro Leu Asn Gly Cys Gly Thr Ar - #g Tyr Lys Phe Glu Asp                       420      - #           425      - #           430                  - - GAT AAA GTC GTC TAT GAA AAC GAA ATA CAT GC - #T CTC TGG ACG GAT TTT         1344                                                                       Asp Lys Val Val Tyr Glu Asn Glu Ile His Al - #a Leu Trp Thr Asp Phe                   435          - #       440          - #       445                      - - CCT CCA AGC AAA ATA TCT AGA GAC AGT GAG TT - #C AGA ATG ACA GTG AAG         1392                                                                       Pro Pro Ser Lys Ile Ser Arg Asp Ser Glu Ph - #e Arg Met Thr Val Lys               450              - #   455              - #   460                          - - TGT TCT TAT AGC AGG AAT GAC ATG CTA CTA AA - #C ATC AAC GTT GAA AGC         1440                                                                       Cys Ser Tyr Ser Arg Asn Asp Met Leu Leu As - #n Ile Asn Val Glu Ser           465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - CTT ACT CCT CCA GTG GCC TCA GTG AAG TTG GG - #T CCA TTT ACC TTG        ATC     1488                                                                    Leu Thr Pro Pro Val Ala Ser Val Lys Leu Gl - #y Pro Phe Thr Leu Ile                          485  - #               490  - #               495              - - CTG CAA AGC TAC CCA GAT AAT TCC TAC CAA CA - #A CCT TAT GGG GAA AAC         1536                                                                       Leu Gln Ser Tyr Pro Asp Asn Ser Tyr Gln Gl - #n Pro Tyr Gly Glu Asn                       500      - #           505      - #           510                  - - GAG TAC CCT CTA GTG AGA TTC CTC CGC CAA CC - #A ATT TAC ATG GAA GTG         1584                                                                       Glu Tyr Pro Leu Val Arg Phe Leu Arg Gln Pr - #o Ile Tyr Met Glu Val                   515          - #       520          - #       525                      - - AGA GTC CTA AAC AGG GAT GAC CCC AAC ATC AA - #G CTG GTC TTA GAT GAC         1632                                                                       Arg Val Leu Asn Arg Asp Asp Pro Asn Ile Ly - #s Leu Val Leu Asp Asp               530              - #   535              - #   540                          - - TGC TGG GCG ACG TCC ACC ATG GAT CCA GAC TC - #T TTC CCC CAG TGG AAC         1680                                                                       Cys Trp Ala Thr Ser Thr Met Asp Pro Asp Se - #r Phe Pro Gln Trp Asn           545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - GTT GTC GTG GAT GGC TGT GCA TAT GAC CTG GA - #C AAC TAC CAG ACC        ACC     1728                                                                    Val Val Val Asp Gly Cys Ala Tyr Asp Leu As - #p Asn Tyr Gln Thr Thr                          565  - #               570  - #               575              - - TTC CAT CCA GTC GGC TCC TCT GTG ACC CAT CC - #T GAT CAC TAT CAG AGG         1776                                                                       Phe His Pro Val Gly Ser Ser Val Thr His Pr - #o Asp His Tyr Gln Arg                       580      - #           585      - #           590                  - - TTT GAC ATG AAG GCT TTT GCC TTT GTA TCA GA - #A GCC CAC GTG CTC TCT         1824                                                                       Phe Asp Met Lys Ala Phe Ala Phe Val Ser Gl - #u Ala His Val Leu Ser                   595          - #       600          - #       605                      - - AGC CTG GTC TAC TTC CAC TGC AGT GCC TTA AT - #C TGT AAT CGA CTC TCC         1872                                                                       Ser Leu Val Tyr Phe His Cys Ser Ala Leu Il - #e Cys Asn Arg Leu Ser               610              - #   615              - #   620                          - - CCT GAC TCC CCA CTG TGT TCT GTG ACC TGC CC - #T GTG TCC TCT AGG CAC         1920                                                                       Pro Asp Ser Pro Leu Cys Ser Val Thr Cys Pr - #o Val Ser Ser Arg His           625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - AGG CGA GCC ACA GGG GCC ACT GAA GCA GAG AA - #A ATG ACA GTC AGC        CTC     1968                                                                    Arg Arg Ala Thr Gly Ala Thr Glu Ala Glu Ly - #s Met Thr Val Ser Leu                          645  - #               650  - #               655              - - CCA GGA CCC ATT CTC CTG TTG TCA GAT GAC TC - #C TCA TTC AGA GGT GTC         2016                                                                       Pro Gly Pro Ile Leu Leu Leu Ser Asp Asp Se - #r Ser Phe Arg Gly Val                       660      - #           665      - #           670                  - - GGC TCA TCT GAT CTA AAA GCA AGT GGG AGC AG - #T GGG GAG AAG AGT AGG         2064                                                                       Gly Ser Ser Asp Leu Lys Ala Ser Gly Ser Se - #r Gly Glu Lys Ser Arg                   675          - #       680          - #       685                      - - AGT GAA ACA GGG GAG GAG GTT GGC TCA CGA GG - #T GCT ATG GAC ACC AAA         2112                                                                       Ser Glu Thr Gly Glu Glu Val Gly Ser Arg Gl - #y Ala Met Asp Thr Lys               690              - #   695              - #   700                          - - GGG CAC AAG ACT GCT GGA GAT GTT GGT TCC AA - #A GCT GTG GCT GCT GTG         2160                                                                       Gly His Lys Thr Ala Gly Asp Val Gly Ser Ly - #s Ala Val Ala Ala Val           705                 7 - #10                 7 - #15                 7 -      #20                                                                              - - GCT GCC TTT GCA GGT GTG GTG GCA ACT CTA GG - #C TTC ATC TAC TAC        CTG     2208                                                                    Ala Ala Phe Ala Gly Val Val Ala Thr Leu Gl - #y Phe Ile Tyr Tyr Leu                          725  - #               730  - #               735              - - TAC GAG AAA AGG ACT GTG TCA AAT CAC TAAATGGGC - #T TCTAAATAAA               2255                                                                       Tyr Glu Lys Arg Thr Val Ser Asn His                                                       740      - #           745                                         - - GCAGTCAAAA T               - #                  - #                      - #     2266                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:43:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 745 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                              - - Met Ala Cys Arg Gln Arg Gly Gly Ser Trp Se - #r Pro Ser Gly Trp Phe        1               5 - #                 10 - #                 15              - - Asn Ala Gly Trp Ser Thr Tyr Arg Ser Ile Se - #r Leu Phe Phe Ala Leu                   20     - #             25     - #             30                  - - Val Thr Ser Gly Asn Ser Ile Asp Val Ser Gl - #n Leu Val Asn Pro Ala               35         - #         40         - #         45                      - - Phe Pro Gly Thr Val Thr Cys Asp Glu Arg Gl - #u Ile Thr Val Glu Phe           50             - #     55             - #     60                          - - Pro Ser Ser Pro Gly Thr Lys Lys Trp His Al - #a Ser Val Val Asp Pro       65                 - # 70                 - # 75                 - # 80       - - Leu Gly Leu Asp Met Pro Asn Cys Thr Tyr Il - #e Leu Asp Pro Glu Lys                       85 - #                 90 - #                 95              - - Leu Thr Leu Arg Ala Thr Tyr Asp Asn Cys Th - #r Arg Arg Val His Gly                  100      - #           105      - #           110                  - - Gly His Gln Met Thr Ile Arg Val Met Asn As - #n Ser Ala Ala Leu Arg              115          - #       120          - #       125                      - - His Gly Ala Val Met Tyr Gln Phe Phe Cys Pr - #o Ala Met Gln Val Glu          130              - #   135              - #   140                          - - Glu Thr Gln Gly Leu Ser Ala Ser Thr Ile Cy - #s Gln Lys Asp Phe Met      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ser Phe Ser Leu Pro Arg Val Phe Ser Gly Le - #u Ala Asp Asp Ser        Lys                                                                                             165  - #               170  - #               175             - - Gly Thr Lys Val Gln Met Gly Trp Ser Ile Gl - #u Val Gly Asp Gly Ala                  180      - #           185      - #           190                  - - Arg Ala Lys Thr Leu Thr Leu Pro Glu Ala Me - #t Lys Glu Gly Phe Ser              195          - #       200          - #       205                      - - Leu Leu Ile Asp Asn His Arg Met Thr Phe Hi - #s Val Pro Phe Asn Ala          210              - #   215              - #   220                          - - Thr Gly Val Thr His Tyr Val Gln Gly Asn Se - #r His Leu Tyr Met Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ser Leu Lys Leu Thr Phe Ile Ser Pro Gly Gl - #n Lys Val Ile Phe        Ser                                                                                             245  - #               250  - #               255             - - Ser Gln Ala Ile Cys Ala Pro Asp Pro Val Th - #r Cys Asn Ala Thr His                  260      - #           265      - #           270                  - - Met Thr Leu Thr Ile Pro Glu Phe Pro Gly Ly - #s Leu Lys Ser Val Ser              275          - #       280          - #       285                      - - Phe Glu Asn Gln Asn Ile Asp Val Ser Gln Le - #u His Asp Asn Gly Ile          290              - #   295              - #   300                          - - Asp Leu Glu Ala Thr Asn Gly Met Lys Leu Hi - #s Phe Ser Lys Thr Leu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Leu Lys Thr Lys Leu Ser Glu Lys Cys Leu Le - #u His Gln Phe Tyr        Leu                                                                                             325  - #               330  - #               335             - - Ala Ser Leu Lys Leu Thr Phe Leu Leu Arg Pr - #o Glu Thr Val Ser Met                  340      - #           345      - #           350                  - - Val Ile Tyr Pro Glu Cys Leu Cys Glu Ser Pr - #o Val Ser Ile Val Thr              355          - #       360          - #       365                      - - Gly Glu Leu Cys Thr Gln Asp Gly Phe Met As - #p Val Glu Val Tyr Ser          370              - #   375              - #   380                          - - Tyr Gln Thr Gln Pro Ala Leu Asp Leu Gly Th - #r Leu Arg Val Gly Asn      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Ser Cys Gln Pro Val Phe Glu Ala Gln Se - #r Gln Gly Leu Val        Arg                                                                                             405  - #               410  - #               415             - - Phe His Ile Pro Leu Asn Gly Cys Gly Thr Ar - #g Tyr Lys Phe Glu Asp                  420      - #           425      - #           430                  - - Asp Lys Val Val Tyr Glu Asn Glu Ile His Al - #a Leu Trp Thr Asp Phe              435          - #       440          - #       445                      - - Pro Pro Ser Lys Ile Ser Arg Asp Ser Glu Ph - #e Arg Met Thr Val Lys          450              - #   455              - #   460                          - - Cys Ser Tyr Ser Arg Asn Asp Met Leu Leu As - #n Ile Asn Val Glu Ser      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Leu Thr Pro Pro Val Ala Ser Val Lys Leu Gl - #y Pro Phe Thr Leu        Ile                                                                                             485  - #               490  - #               495             - - Leu Gln Ser Tyr Pro Asp Asn Ser Tyr Gln Gl - #n Pro Tyr Gly Glu Asn                  500      - #           505      - #           510                  - - Glu Tyr Pro Leu Val Arg Phe Leu Arg Gln Pr - #o Ile Tyr Met Glu Val              515          - #       520          - #       525                      - - Arg Val Leu Asn Arg Asp Asp Pro Asn Ile Ly - #s Leu Val Leu Asp Asp          530              - #   535              - #   540                          - - Cys Trp Ala Thr Ser Thr Met Asp Pro Asp Se - #r Phe Pro Gln Trp Asn      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Val Val Val Asp Gly Cys Ala Tyr Asp Leu As - #p Asn Tyr Gln Thr        Thr                                                                                             565  - #               570  - #               575             - - Phe His Pro Val Gly Ser Ser Val Thr His Pr - #o Asp His Tyr Gln Arg                  580      - #           585      - #           590                  - - Phe Asp Met Lys Ala Phe Ala Phe Val Ser Gl - #u Ala His Val Leu Ser              595          - #       600          - #       605                      - - Ser Leu Val Tyr Phe His Cys Ser Ala Leu Il - #e Cys Asn Arg Leu Ser          610              - #   615              - #   620                          - - Pro Asp Ser Pro Leu Cys Ser Val Thr Cys Pr - #o Val Ser Ser Arg His      625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - Arg Arg Ala Thr Gly Ala Thr Glu Ala Glu Ly - #s Met Thr Val Ser        Leu                                                                                             645  - #               650  - #               655             - - Pro Gly Pro Ile Leu Leu Leu Ser Asp Asp Se - #r Ser Phe Arg Gly Val                  660      - #           665      - #           670                  - - Gly Ser Ser Asp Leu Lys Ala Ser Gly Ser Se - #r Gly Glu Lys Ser Arg              675          - #       680          - #       685                      - - Ser Glu Thr Gly Glu Glu Val Gly Ser Arg Gl - #y Ala Met Asp Thr Lys          690              - #   695              - #   700                          - - Gly His Lys Thr Ala Gly Asp Val Gly Ser Ly - #s Ala Val Ala Ala Val      705                 7 - #10                 7 - #15                 7 -      #20                                                                              - - Ala Ala Phe Ala Gly Val Val Ala Thr Leu Gl - #y Phe Ile Tyr Tyr        Leu                                                                                             725  - #               730  - #               735             - - Tyr Glu Lys Arg Thr Val Ser Asn His                                                  740      - #           745                                         - -  - - (2) INFORMATION FOR SEQ ID NO:44:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 560 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 15..506                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                              - - GAATTCGCGG CCGC TCC TCT GTG ACC CAT CCT GAT C - #AC TAT CAG AGG TTT           50                                                                                       Ser S - #er Val Thr His Pro Asp His Tyr Gln Arg Ph - #e                         - #1               5   - #               10                  - - GAC ATG AAG GCT TTT GCC TTT GTA TCA GAG GC - #C CAT GTG CTC TCT AGC           98                                                                       Asp Met Lys Ala Phe Ala Phe Val Ser Glu Al - #a His Val Leu Ser Ser                    15         - #         20         - #         25                      - - CTG GTC TAC TTC CAC TGC AGT GCC TTA ATC TG - #C AAT CGA CTC TCT CCA          146                                                                       Leu Val Tyr Phe His Cys Ser Ala Leu Ile Cy - #s Asn Arg Leu Ser Pro                30             - #     35             - #     40                          - - GAC TCC CCT CTG TGT TCT GTG ACC TGC CCT GT - #G TCA TCT AGG CAC AGG          194                                                                       Asp Ser Pro Leu Cys Ser Val Thr Cys Pro Va - #l Ser Ser Arg His Arg            45                 - # 50                 - # 55                 - # 60       - - CGA GCC ACA GGG GCC ACT GAA GCA GAG AAA AT - #G ACA GTC AGC CTC CCA          242                                                                       Arg Ala Thr Gly Ala Thr Glu Ala Glu Lys Me - #t Thr Val Ser Leu Pro                            65 - #                 70 - #                 75              - - GGA CCC ATT CTC CTG TTG TCA GAC GAC TCC TC - #A TTC AGA GGT GTT GGC          290                                                                       Gly Pro Ile Leu Leu Leu Ser Asp Asp Ser Se - #r Phe Arg Gly Val Gly                        80     - #             85     - #             90                  - - TCA TCT GAT CTA AAA GCA AGT GGG AGC AGT GG - #G GAG AAC AGT AGG AGC          338                                                                       Ser Ser Asp Leu Lys Ala Ser Gly Ser Ser Gl - #y Glu Asn Ser Arg Ser                    95         - #        100         - #        105                      - - GAA ACA GGG GAG GAG GTT GGC TCA CGA GAT GT - #T ATG GAC ACC AAA GGG          386                                                                       Glu Thr Gly Glu Glu Val Gly Ser Arg Asp Va - #l Met Asp Thr Lys Gly               110              - #   115              - #   120                          - - CAC AGG ACT GCT GGA GAT GTT GGT TCC AAA GC - #T GTG GCT GCT GTG GCT          434                                                                       His Arg Thr Ala Gly Asp Val Gly Ser Lys Al - #a Val Ala Ala Val Ala           125                 1 - #30                 1 - #35                 1 -      #40                                                                              - - GCC TTG GCA GGT GTG GTG GCA ACT CTA GGC TT - #C ATC TGT TAC CTG        TAT      482                                                                    Ala Leu Ala Gly Val Val Ala Thr Leu Gly Ph - #e Ile Cys Tyr Leu Tyr                          145  - #               150  - #               155              - - AAG AAA AGG ACT GTG TCA AAT CAC TAAATGGGCT TC - #TAAATAAA GCAGTCAAAA         536                                                                       Lys Lys Arg Thr Val Ser Asn His                                                           160                                                                - - TAAAAAAAAA GCGGCCGCGA ATTC          - #                  - #                   560                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:45:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 164 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                              - - Ser Ser Val Thr His Pro Asp His Tyr Gln Ar - #g Phe Asp Met Lys Ala        1               5 - #                 10 - #                 15              - - Phe Ala Phe Val Ser Glu Ala His Val Leu Se - #r Ser Leu Val Tyr Phe                   20     - #             25     - #             30                  - - His Cys Ser Ala Leu Ile Cys Asn Arg Leu Se - #r Pro Asp Ser Pro Leu               35         - #         40         - #         45                      - - Cys Ser Val Thr Cys Pro Val Ser Ser Arg Hi - #s Arg Arg Ala Thr Gly           50             - #     55             - #     60                          - - Ala Thr Glu Ala Glu Lys Met Thr Val Ser Le - #u Pro Gly Pro Ile Leu       65                 - # 70                 - # 75                 - # 80       - - Leu Leu Ser Asp Asp Ser Ser Phe Arg Gly Va - #l Gly Ser Ser Asp Leu                       85 - #                 90 - #                 95              - - Lys Ala Ser Gly Ser Ser Gly Glu Asn Ser Ar - #g Ser Glu Thr Gly Glu                  100      - #           105      - #           110                  - - Glu Val Gly Ser Arg Asp Val Met Asp Thr Ly - #s Gly His Arg Thr Ala              115          - #       120          - #       125                      - - Gly Asp Val Gly Ser Lys Ala Val Ala Ala Va - #l Ala Ala Leu Ala Gly          130              - #   135              - #   140                          - - Val Val Ala Thr Leu Gly Phe Ile Cys Tyr Le - #u Tyr Lys Lys Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Val Ser Asn His                                                           - -  - - (2) INFORMATION FOR SEQ ID NO:46:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 866 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 12..821                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                              - - GAATTCGCGG C CGC CGT GGC TCT GTC ACT CGT GAC - #AGC ATC TTC AGG        CTC      50                                                                                  Arg Arg Gly - #Ser Val Thr Arg Asp Ser Ile Phe Arg Leu                         1   - #            5      - #            10                     - - CAT GTC AGC TGC AGC TAC TCA GTA AGT AGC AA - #C TCT CTC CCA ATC AAG           98                                                                       His Val Ser Cys Ser Tyr Ser Val Ser Ser As - #n Ser Leu Pro Ile Lys                15             - #     20             - #     25                          - - GTC CAG GTT TTT ACT CTC CCA CCA CCC TTT CC - #T GAG ACC CAG CCT GGA          146                                                                       Val Gln Val Phe Thr Leu Pro Pro Pro Phe Pr - #o Glu Thr Gln Pro Gly            30                 - # 35                 - # 40                 - # 45       - - CCC CTC ACT CTG GAA CTT CAG ATT GCC AAA GA - #T AAA AAC TAT GGC TCC          194                                                                       Pro Leu Thr Leu Glu Leu Gln Ile Ala Lys As - #p Lys Asn Tyr Gly Ser                            50 - #                 55 - #                 60              - - TAC TAT GGT GTT GGT GAC TAC CCC GTG GTG AA - #G TTG CTT CGG GAT CCC          242                                                                       Tyr Tyr Gly Val Gly Asp Tyr Pro Val Val Ly - #s Leu Leu Arg Asp Pro                        65     - #             70     - #             75                  - - ATC TAT GTG GAG GTC TCC ATC CTT CAC AGA AC - #A GAC CCC TCC CTG GGG          290                                                                       Ile Tyr Val Glu Val Ser Ile Leu His Arg Th - #r Asp Pro Ser Leu Gly                    80         - #         85         - #         90                      - - CTG CTC CTA CAT CAG TGT TGG GCA ACA CCC AG - #C ACA GAC CCA CTG AGT          338                                                                       Leu Leu Leu His Gln Cys Trp Ala Thr Pro Se - #r Thr Asp Pro Leu Ser                95             - #    100             - #    105                          - - CAG CCA CAG TGG CCC ATC CTG GTA AAG GGC TG - #C CCC TAC ATT GGA GAC          386                                                                       Gln Pro Gln Trp Pro Ile Leu Val Lys Gly Cy - #s Pro Tyr Ile Gly Asp           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - AAC TAT CAG ACC CAG CTG ATC CCT GTC CAG AA - #A GCC TTG GAT CTT        CCA      434                                                                    Asn Tyr Gln Thr Gln Leu Ile Pro Val Gln Ly - #s Ala Leu Asp Leu Pro                          130  - #               135  - #               140              - - TTT CCC TCT CAC TAC CAG CGC TTC AGC ATC TT - #C ACC TTC AGC TTT GTG          482                                                                       Phe Pro Ser His Tyr Gln Arg Phe Ser Ile Ph - #e Thr Phe Ser Phe Val                       145      - #           150      - #           155                  - - GAC CCT ACA GCG GAG AAA CAG GCC CTC AGG GG - #A CCG GTG CAT CTG CAC          530                                                                       Asp Pro Thr Ala Glu Lys Gln Ala Leu Arg Gl - #y Pro Val His Leu His                   160          - #       165          - #       170                      - - TGC AGT GTG TCA GTC TGC CAG CCT GCT GAG AC - #A CCA TCC TGT GCG GTA          578                                                                       Cys Ser Val Ser Val Cys Gln Pro Ala Glu Th - #r Pro Ser Cys Ala Val               175              - #   180              - #   185                          - - ACC TGT CCT GAT CTC AGT CGA AGA AAT TCA GG - #C ACC ATT TTT CAG AAC          626                                                                       Thr Cys Pro Asp Leu Ser Arg Arg Asn Ser Gl - #y Thr Ile Phe Gln Asn           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - ACT ACT GCT AGT GTT TCT AGC AAA GGC CCC AT - #G ATT CTA CTC CAA        GCC      674                                                                    Thr Thr Ala Ser Val Ser Ser Lys Gly Pro Me - #t Ile Leu Leu Gln Ala                          210  - #               215  - #               220              - - ACT AAG GAC CCT CCA GAA AAG CTC CGT GCT CC - #T GTA GAC TCA AAA GTT          722                                                                       Thr Lys Asp Pro Pro Glu Lys Leu Arg Ala Pr - #o Val Asp Ser Lys Val                       225      - #           230      - #           235                  - - CTG TGG GTG GCA GGC CTT TCT GGG ACC TTA AT - #C CTT GGA GGC TTA GTA          770                                                                       Leu Trp Val Ala Gly Leu Ser Gly Thr Leu Il - #e Leu Gly Gly Leu Val                   240          - #       245          - #       250                      - - GTA TCC TAC TTG GCT ATC AAA CAG CTG AAT TG - #T CCA GAC CAA ACA TGT          818                                                                       Val Ser Tyr Leu Ala Ile Lys Gln Leu Asn Cy - #s Pro Asp Gln Thr Cys               255              - #   260              - #   265                          - - CAA TAAAACCAGA CTGTACTCCC AAAAAAAAAA AGCGGCCGCG AATTC - #                   866                                                                        Gln                                                                            - - 270                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:47:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 270 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                              - - Arg Arg Gly Ser Val Thr Arg Asp Ser Ile Ph - #e Arg Leu His Val Ser        1               5 - #                 10 - #                 15              - - Cys Ser Tyr Ser Val Ser Ser Asn Ser Leu Pr - #o Ile Lys Val Gln Val                   20     - #             25     - #             30                  - - Phe Thr Leu Pro Pro Pro Phe Pro Glu Thr Gl - #n Pro Gly Pro Leu Thr               35         - #         40         - #         45                      - - Leu Glu Leu Gln Ile Ala Lys Asp Lys Asn Ty - #r Gly Ser Tyr Tyr Gly           50             - #     55             - #     60                          - - Val Gly Asp Tyr Pro Val Val Lys Leu Leu Ar - #g Asp Pro Ile Tyr Val       65                 - # 70                 - # 75                 - # 80       - - Glu Val Ser Ile Leu His Arg Thr Asp Pro Se - #r Leu Gly Leu Leu Leu                       85 - #                 90 - #                 95              - - His Gln Cys Trp Ala Thr Pro Ser Thr Asp Pr - #o Leu Ser Gln Pro Gln                  100      - #           105      - #           110                  - - Trp Pro Ile Leu Val Lys Gly Cys Pro Tyr Il - #e Gly Asp Asn Tyr Gln              115          - #       120          - #       125                      - - Thr Gln Leu Ile Pro Val Gln Lys Ala Leu As - #p Leu Pro Phe Pro Ser          130              - #   135              - #   140                          - - His Tyr Gln Arg Phe Ser Ile Phe Thr Phe Se - #r Phe Val Asp Pro Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ala Glu Lys Gln Ala Leu Arg Gly Pro Val Hi - #s Leu His Cys Ser        Val                                                                                             165  - #               170  - #               175             - - Ser Val Cys Gln Pro Ala Glu Thr Pro Ser Cy - #s Ala Val Thr Cys Pro                  180      - #           185      - #           190                  - - Asp Leu Ser Arg Arg Asn Ser Gly Thr Ile Ph - #e Gln Asn Thr Thr Ala              195          - #       200          - #       205                      - - Ser Val Ser Ser Lys Gly Pro Met Ile Leu Le - #u Gln Ala Thr Lys Asp          210              - #   215              - #   220                          - - Pro Pro Glu Lys Leu Arg Ala Pro Val Asp Se - #r Lys Val Leu Trp Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ala Gly Leu Ser Gly Thr Leu Ile Leu Gly Gl - #y Leu Val Val Ser        Tyr                                                                                             245  - #               250  - #               255             - - Leu Ala Ile Lys Gln Leu Asn Cys Pro Asp Gl - #n Thr Cys Gln                          260      - #           265      - #           270                  - -  - - (2) INFORMATION FOR SEQ ID NO:48:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 722 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 15..683                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                              - - GAATTCGCGG CCGC ATC CAC ACT GGC AGC CAC GTG C - #CA CTG CGG TTG TTT           50                                                                                       Ile H - #is Thr Gly Ser His Val Pro Leu Arg Leu Ph - #e                         - #1               5   - #               10                  - - GTG GAC CAC TGC GTG GCC ACA CCA ACA CCA GA - #C CAG AAT GCC TCC CCT           98                                                                       Val Asp His Cys Val Ala Thr Pro Thr Pro As - #p Gln Asn Ala Ser Pro                    15         - #         20         - #         25                      - - TAT CAC ACC ATC GTG GAC TTC CAT GGC TGT CT - #T GTC GAT GGT CTC ACT          146                                                                       Tyr His Thr Ile Val Asp Phe His Gly Cys Le - #u Val Asp Gly Leu Thr                30             - #     35             - #     40                          - - GAT GCC TCT TCT GCG TTC AAA GTT CCT CGA CC - #C GGG CCA GAT ACA CTC          194                                                                       Asp Ala Ser Ser Ala Phe Lys Val Pro Arg Pr - #o Gly Pro Asp Thr Leu            45                 - # 50                 - # 55                 - # 60       - - CAG TTC ACA GTG GAT GTC TTC CAC TTT GCT AA - #T GAC TCC AGA AAC ATG          242                                                                       Gln Phe Thr Val Asp Val Phe His Phe Ala As - #n Asp Ser Arg Asn Met                            65 - #                 70 - #                 75              - - ATA TAC ATC ACC TGC CAC CTG AAG GCC ATC CC - #A GCT GAG CAG GAA CCA          290                                                                       Ile Tyr Ile Thr Cys His Leu Lys Ala Ile Pr - #o Ala Glu Gln Glu Pro                        80     - #             85     - #             90                  - - GAC GAA CTC AAC AAA GCC TGT TCC TTC AGC AA - #G TCT TCC AAC AGC TGG          338                                                                       Asp Glu Leu Asn Lys Ala Cys Ser Phe Ser Ly - #s Ser Ser Asn Ser Trp                    95         - #        100         - #        105                      - - TTC CCA GTG GAA GGC CCA GCT GAC ATC TGT CA - #A TGC TGT AGC AAG GGT          386                                                                       Phe Pro Val Glu Gly Pro Ala Asp Ile Cys Gl - #n Cys Cys Ser Lys Gly               110              - #   115              - #   120                          - - GAC TGT GGC ACT CCA AGC CAT TCC AGG AGG CA - #G CCC CAT GTC GTG AGC          434                                                                       Asp Cys Gly Thr Pro Ser His Ser Arg Arg Gl - #n Pro His Val Val Ser           125                 1 - #30                 1 - #35                 1 -      #40                                                                              - - CAG TGG TCC AGG TCT GCT TCT CGT AAC CGC AG - #G CAT GTG ACA GAA        GAA      482                                                                    Gln Trp Ser Arg Ser Ala Ser Arg Asn Arg Ar - #g His Val Thr Glu Glu                          145  - #               150  - #               155              - - GCA GAT ATC ACC GTG GGG CCA CTG ATC TTC CT - #G GAC AGG AGT GCT GAC          530                                                                       Ala Asp Ile Thr Val Gly Pro Leu Ile Phe Le - #u Asp Arg Ser Ala Asp                       160      - #           165      - #           170                  - - TAT GAA GTA GAA CAG TGG GCC TTG CCG ACT GA - #C ACC TCC GTG CTG CTG          578                                                                       Tyr Glu Val Glu Gln Trp Ala Leu Pro Thr As - #p Thr Ser Val Leu Leu                   175          - #       180          - #       185                      - - CTG GGC ATA GGC CTG GCC GTG GTG GCA TCT CT - #G ACT CTG ACC GCT GTT          626                                                                       Leu Gly Ile Gly Leu Ala Val Val Ala Ser Le - #u Thr Leu Thr Ala Val               190              - #   195              - #   200                          - - ATC CTG ATT TTC ACC AGG AGG TGG CGC ACT GC - #C TCC CGC CCT GTG TCT          674                                                                       Ile Leu Ile Phe Thr Arg Arg Trp Arg Thr Al - #a Ser Arg Pro Val Ser           205                 2 - #10                 2 - #15                 2 -      #20                                                                              - - GTT TCC CAA TAAAAGAAGA AAGCAGTAAA AAAAAGCGGC CGCGAATTC - #                   722                                                                      Val Ser Gln                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:49:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 223 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                              - - Ile His Thr Gly Ser His Val Pro Leu Arg Le - #u Phe Val Asp His Cys        1               5 - #                 10 - #                 15              - - Val Ala Thr Pro Thr Pro Asp Gln Asn Ala Se - #r Pro Tyr His Thr Ile                   20     - #             25     - #             30                  - - Val Asp Phe His Gly Cys Leu Val Asp Gly Le - #u Thr Asp Ala Ser Ser               35         - #         40         - #         45                      - - Ala Phe Lys Val Pro Arg Pro Gly Pro Asp Th - #r Leu Gln Phe Thr Val           50             - #     55             - #     60                          - - Asp Val Phe His Phe Ala Asn Asp Ser Arg As - #n Met Ile Tyr Ile Thr       65                 - # 70                 - # 75                 - # 80       - - Cys His Leu Lys Ala Ile Pro Ala Glu Gln Gl - #u Pro Asp Glu Leu Asn                       85 - #                 90 - #                 95              - - Lys Ala Cys Ser Phe Ser Lys Ser Ser Asn Se - #r Trp Phe Pro Val Glu                  100      - #           105      - #           110                  - - Gly Pro Ala Asp Ile Cys Gln Cys Cys Ser Ly - #s Gly Asp Cys Gly Thr              115          - #       120          - #       125                      - - Pro Ser His Ser Arg Arg Gln Pro His Val Va - #l Ser Gln Trp Ser Arg          130              - #   135              - #   140                          - - Ser Ala Ser Arg Asn Arg Arg His Val Thr Gl - #u Glu Ala Asp Ile Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Val Gly Pro Leu Ile Phe Leu Asp Arg Ser Al - #a Asp Tyr Glu Val        Glu                                                                                             165  - #               170  - #               175             - - Gln Trp Ala Leu Pro Thr Asp Thr Ser Val Le - #u Leu Leu Gly Ile Gly                  180      - #           185      - #           190                  - - Leu Ala Val Val Ala Ser Leu Thr Leu Thr Al - #a Val Ile Leu Ile Phe              195          - #       200          - #       205                      - - Thr Arg Arg Trp Arg Thr Ala Ser Arg Pro Va - #l Ser Val Ser Gln              210              - #   215              - #   220                          - -  - - (2) INFORMATION FOR SEQ ID NO:50:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 36 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                              - - CGCCCTTCCC AGCAACTGCA CCATCACCAC CATGGG      - #                  -     #       36                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:51:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 45 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                              - - GATCCCCATG GTGGTGGTGA TGGTGCAGTT GCTGGGAAGG GCGAT   - #                      - #45                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:52:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                              - - GATCCCTCGA GCCACCATCA CCACCATCAT G        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:53:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                              - - AATTCATGAT GGTGGTGATG GTGGCTCGAG G        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:54:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                              - - CCCGGATCCG CAGACCATCT GGCCAACTGA G        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:55:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                              - - GCGCTCGAGG GCATATGGCT GCCAGTGTG         - #                  - #                29                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:56:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 36 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                              - - CGCGCTAGCA GATCTATGGC GCCGAGCTGG AGGTTC      - #                  -     #       36                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:57:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 49 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                              - - CGCGGATCCT ATTAATGGTG GTGATGGTGG TGACTAGTGG ACCCTTCCA  - #                   49                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:58:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 39 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                              - - CCCGCTAGCA GATCTATGGG GCTGAGCTAT GGAATTTTC      - #                      - #    39                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:59:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 34 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                              - - CGCACTAGTT GACCCCTCTA TACCATGATC ACTA       - #                  -      #        34                                                                   __________________________________________________________________________

We claim:
 1. An isolated DNA encoding a ZPA polypeptide, the DNA beingselected from the group consisting of DNA set out in SEQ ID NOS:. 9, 13,and 19;or a DNA encoding an immunologically active fragment of the ZPApolypeptide encoded by a DNA selected from the group consisting of SEQID NOS:. 9, 13, and
 19. 2. A DNA encoding a ZPA polypeptide saidpolypeptide comprising an amino acid sequence selected from the groupconsisting of SEQ ID NOS:. 10, 14, and 20;or a DNA encoding animmunologically active fragment of said ZPA polypeptide selected fromthe group consisting of SEQ ID NOS: 10, 14, and
 20. 3. A vector whichcomprises a DNA according to claim 1 wherein said DNA comprises the DNAset out in SEQ ID NO.
 9. 4. A vector which comprises a DNA according toclaim 1 wherein said DNA comprises the DNA set out in SEQ ID NO.
 13. 5.A vector which comprises a DNA according to claim 1 wherein said DNAcomprises the DNA set out in SEQ ID NO.
 19. 6. An isolated DNA encodinga ZPA polypeptide comprising DNA consisting of the set forth in SEQ IDNO.
 1. 7. A DNA encoding a ZPA polypeptide said polypeptide comprisingan amino acid sequence according to SEQ ID NO.
 2. 8. A vector whichcomprises a DNA according to claim
 6. 9. A procaryotic or eucaryotichost cell stably transformed or transfected with a vector according toclaims 3, 4, or
 5. 10. A process for the production of a mammalian ZPApolypeptide or an immunologically active fragment thereof, said processcomprising:(a) growing, under suitable nutrient conditions, procaryoticor eucaryotic host cells according to claim 9; and (b) isolating the ZPApolypeptide or immunologically active fragment of ZPA producedtherefrom.
 11. A procaryotic or eucaryotic host cell stably transformedor transfected with a vector according to claim
 8. 12. A process for theproduction of a mammalian ZPA polypeptide said process comprising:(a)growing, under suitable nutrient conditions, procaryotic or eucaryotichost cells according to claim 11; and (b) isolating the ZPA polypeptideproduced therefrom.
 13. A polynucleotide comprising a nucleotidesequence as set out in SEQ ID NO.
 1. 14. A polynucleotide comprising anucleotide sequence as set out in SEQ ID NO.
 9. 15. A polynucleotidecomprising a nucleotide sequence as set out in SEQ ID NO.
 13. 16. Apolynucleotide comprising a nucleotide sequence as set out in SEQ ID NO.19.
 17. A polynucleotide comprising a nucleotide sequence as set out inSEQ ID NO.
 44. 18. An isolated DNA encoding a ZPA polypeptide, the DNAset out in SEQ ID NO.
 44. 19. A DNA encoding a ZPA polypeptide, saidpolypeptide comprising an amino acid sequence set out in SEQ ID NO. 45.20. A vector which comprises a DNA according to claim
 18. 21. Aproaryotic or eucaryotic host cell stably transformed or transfectedwith a vector according to claim
 20. 22. A process for the production ofa mammalian ZPA polypeptide, said process comprising:(a) growing, undersuitable nutrient conditions, procaryotic or eucaryotic host cellsaccording to claim 21; and b) isolating the ZPA polypeptide producedtherefrom.